The present research examines the conditions under which educating non-stigmatized individuals about the experiences of members of stigmatized groups leads to paternalistic or more respectful views of the target. We propose that when these efforts ask members of non-stigmatized groups to focus only on the difficulties experienced by stigmatized targets, they will lead to more paternalistic views of targets because they portray targets as being in need of help. In contrast, we propose that when these efforts take a broader focus on stigmatized targets and include their resilience in the face of their difficulties, they will lead to more respectful views of targets. Four studies supported these predictions. Across studies, White participants who focused only on a Black target’s difficulties subsequently perceived the target as more helpless and less competent than controls. Participants who focused on the target’s resilience in the face of difficulties perceived him as more competent. Group-based inequities are an unfortunate reality in modern society. Whereas members of non-stigmatized or majority groups  are afforded privileges and benefits from their group membership, members of stigmatized groups often face considerable disadvantages and marginalization. In the context of such marginalization, intergroup interactions can be fraught with misunderstanding, tension, and ultimately less satisfaction for interaction partners .In recent years, some have argued  that oneTo illustrate why this might be the case, consider as an example a White, middle-class teacher who is attempting to understand the experiences of a struggling student; the student’s grades are lower than they could be, but she also knows that this student comes from a working-class, low-income family and that there are negative stereotypes characterizing his group as intellectually inferior. One way that this teacher could approach the student would be to think about how difficult the student’s plight must be – about how she might feel if she were faced with such hardship. Feeling sorry for the student, the teacher might lower her standards for him: assigning easier problem sets, or calling on him fewer times in class. Although this strategy may lead the teacher to feel sympathy for the student, and perhaps even to like the student more, it does not allow the teacher to appreciate his strengths. By lowering the teacher’s expectations for the student, this approach to understanding the student’s experiences could ultimately prevent him from fulfilling his full academic potential.An alternative way the teacher could attempt to understand the student’s experiences would be to focus on the student’s strengths and resilience in the face of the barriers he faces. With a newfound appreciation of the student and his strengths, the teacher may choose to provide support and encouragement that would allow the student to improve without lowering her standards for him. Although the differences between these two ways of approaching stigma may seem subtle, they could lead to very different outcomes for both the teacher and the student. Following from these possibilities, in the present research we propose two different ways of thinking about societal stigmatization and suggest that these two approaches will have different consequences for perceptions of stigmatized groups.difficulties-focused approach, which is characterized by a narrow focus on the struggles and difficulties that someone experiences as a result of stigma. We hypothesize that this approach will cause people to view targets as helpless victims, rather than as competent individuals deserving of respect. In contrast, one could take what we refer to as a resilience-focused approach to stigma, which is characterized by a focus on a person’s resilience in the face of their difficulties. We suggest that this way of thinking about stigma will lead to increased appreciation of targets’ competencies, and ultimately increased respect for targets.Although there are many ways that one can think about stigma, we propose an important distinction between two such ways in particular. The first we refer to as a augmentation principle, learning that another person has acted in the face of a constraint should lead people to make stronger inferences about the internal causes of the action than if the constraint were not present  participated in the study in exchange for payment. We conducted a sensitivity power analysis with G*Power  first read an article about stereotype threat . The artdifficulties-focus condition were further instructed to “try to imagine how much more difficult his experiences are than the typical student, and the kind of help he might need in such situations”; whereas participants in the resilience-focus condition were instructed to “try to imagine how much stronger he will become from these difficult experiences, and what he could teach you about dealing with challenging situations.” Participants in the control condition received no such further instructions.After reading their respective articles, participants completed a perspective-taking exercise that was modeled after empathy interventions used in previous research . In all All participants were then asked to write one sentence about how they might feel if they were Tyrone in five different scenarios. In the difficulties condition, these scenarios focused on the struggles and difficulties that Tyrone faces as a result of being a member of a negatively stereotyped group. In the resilience condition, the scenarios focused on Tyrone’s resilience in the face of his difficulties. For instance, one scenario in the difficulties condition asked participants to write about how Tyrone might feel when he “meets with his English professor to talk about his term paper, and how he might worry that the professor might judge his abilities in the light of negative stereotypes about Black people”; whereas in the resilience condition they were asked to write about how he might feel when he “meets with his English professor to talk about his term paper, and what strategies he uses to get the most out of the feedback despite the professor’s potential doubts about his ability.” The control condition included scenarios that were identical to those in the experimental conditions, but without the focus on difficulties or resilience. For instance, the analogous control scenario was: “meets with his English professor to talk about his term paper.”victim, incapable, in need of help, weak, needs support; α = .77), and those related to competence . Although these measures did not come from a previously validated scale, they showed reasonable internal consistency and provided a face valid measurement of helplessness and competence, respectively.The primary dependent measures consisted of participants’ perceptions of the extent to which the target possessed traits related to helplessness and competence. Each trait was rated on a five-point scale . Two composites were formed by averaging participants’ ratings of the traits related to helplessness  = 6.99, p = .001. Participants in the difficulties condition  rated the target as more helpless than did participants in the control condition  and the resilience condition  = 2.75, p = .007, d = 0.55). In contrast, participants in the resilience condition viewed the target as no more helpless than did those in the control condition, t(143) = 0.86, p = .39, d = 0.16.Consistent with our predictions, focus condition affected ratings of helplessness, F = 8.16, p < .001. Participants in the difficulties condition  rated the target as less competent than did those in the control condition  and those in the resilience condition . Participants in the resilience condition rated the target as marginally more competent than did those in the control condition, t(143) = 1.81, p = .072, d = 0.38.Focus condition also influenced ratings of competence, These findings provide preliminary evidence that the two ways of approaching stigma (i.e. the difficulties-focused approach vs. the resilience-focused approach) have different consequences for Whites’ perceptions of stigmatized groups. Indeed, participants who focused only on the target’s difficulties and struggles perceived him as more helpless and less competent than controls. In contrast, participants who focused on the target’s resilience perceived him as more competent than controls.A limitation of Study 1, however, is that participants in the experimental conditions read an article about stereotype threat prior to the perspective-taking activity. It is therefore unclear whether learning about stereotype threat is necessary for the observed effects to emerge, or if the differences between the two focus conditions would emerge with the perspective-taking exercise alone. Thus, in Study 2 we removed the stereotype threat article from the focus manipulation to provide a more straightforward test of our model.https://osf.io/q72pz/?view_only=f88f040dec0244f28ba5787f2271693e). As in Study 1, Study 2 consisted of a three-cell experimental design in which participants took the perspective of a negatively stereotyped target while focusing on his difficulties or his resilience, or completed a control perspective-taking task. Consistent with the findings of Study 1, we predicted that focusing on the target’s difficulties would lead Whites to view the Black target as more helpless and less competent. In contrast, we predicted that participants who focused on the target’s resilience would show no increase in their perceptions of the target’s helplessness and instead perceive him as more competent.In Study 2, we sought to replicate the primary findings of the previous studies using a cleaner manipulation . In addition, we pre-registered our hypotheses and analysis plan prior to collection of the data on the Open Science Framework  participated in a study on the “Development of Reading Materials” in exchange for a cash payment. As in Study 1, we conducted sensitivity analyses for the focal contrasts using G*Power  and competence (α = .84).Our first pre-registered prediction was that participants in the difficulties condition would view the target as more helpless relative to both the control condition and the resilience condition. Our second pre-registered prediction was that participants in the difficulties condition would perceive the target as less competent (vs. control), whereas participants in the resilience condition would perceive the target as more competent (vs. control). We tested these predictions using planned comparisons with two-tailed tests.F = 14.09, p < .001. Participants in the difficulties condition viewed the target as more helpless  than did those in both the control condition , and the resilience condition . Those in the resilience condition did not significantly differ from those in the control condition, t[172] = 0.86, p = .39, d = 0.16.As predicted, we found that focus condition significantly affected ratings of helplessness, F = 9.54, p < .001. Participants in the difficulties condition  rated the target as less competent than did those in the control condition  and the resilience condition . Participants in the resilience condition rated the target as more competent than did those in the control condition, t(172) = 2.23, p = .027, d = 0.46.Focus condition also influenced participants’ ratings of the target’s competence, These findings provide additional evidence that the two ways of approaching stigma differentially affect Whites’ perceptions of stigmatized groups. Participants who focused on the Black target’s difficulties viewed him as more helpless and less competent than controls. In contrast, participants who focused on the target’s resilience in the face of his difficulties perceived the target as more competent than controls. Thus, this study replicates the findings of Study 1 and suggests that learning about stereotype threat prior to the perspective-taking exercise is not necessary for the effects of focus condition to emerge.construals of the scenarios, such that participants would construe the target’s experiences in terms of either the difficulties he faces or his resilience in the face of his difficulties. Our predictions, stopping rule, and analysis plan were pre-registered on the Open Science Framework (https://osf.io/q72pz/?view_only=0871d485aede4423aba52ee60d98222b).Studies 1 and 2 suggest that the two ways of approaching stigma have different consequences for peoples’ perceptions of stigmatized targets. A limitation of these studies, however, is that there were subtle differences in scenarios across the experimental conditions. For instance, one scenario in the difficulties condition was “Takes a difficult midterm test while under extra pressure to prove that the negative stereotype about Black people’s abilities isn’t true,” whereas the analogous scenario in the resilience condition was “Takes a difficult midterm test, and what strategies he has learned to use to reduce any anxiety he feels when taking this test.” Thus, although we tried to hold extraneous factors constant, it is possible that our manipulations were confounded with other differences in the content of the scenarios. Moreover, given that the language used in the manipulation instructions  was similar to some of the items used in the measures of helplessness and competence , one might argue that the findings of Studies 1 and 2 were driven by demand effects. Study 3 addresses these issues by holding the scenarios constant across conditions and manipulating participants’ Mage = 37.98, SD = 11.24) were recruited to participate in the study via TurkPrime.9 A sensitivity analysis revealed that Study 3 was adequately powered to detect an effect size of d = 0.29.Three hundred eighty-four White American MTurk workers , but holding the content across the two conditions constant.construals of the scenarios. To do so, we first asked participants to complete a series of reading and writing activities that varied in terms of whether they focused on the difficulties that negatively stereotyped individuals face or on their resilience in the face of their difficulties .Participants were randomly assigned to a difficulties-focus condition or a resilience-focus condition. Whereas in the previous studies we manipulated participants’ focus by varying the content of the scenarios themselves, in the present study we included the same scenarios across both conditions but manipulated participants’ In both conditions, participants first read an adapted version of the article about stereotype threat that was used in Study 1. Both conditions included a basic overview of stereotype threat, a thought experiment in which participants imagined experiencing stereotype threat, and stories of two individuals’  experiences with stereotype threat. We included examples of female engineers  because our goal was to manipulate participants’ construals of experiences of stigma in a broad sense, beyond just a single Black male’s experiences of stigma.In the difficulties condition the content focused on the difficulties associated with experiencing stereotype threat, whereas in the resilience condition the content focused on the target’s resilience in the face of stereotype threat. For instance, in the difficulties condition participants were told that, because of stereotype threat, Maria  “has to deal with this extra pressure to perform well and to not conform to this stereotype” and that “all of this makes the engineering program a very difficult and stressful situation for Maria.” The resilience condition included this content as well but then went on to say that Maria “uses her strengths and skills to manage the extra stress and pressure.”Both conditions then asked participants to complete a perspective-taking exercise similar to the one used in Study 1, but with a female engineer named “Katherine Parker” as the target of the exercise. The goal of this perspective-taking activity was not to manipulate people’s perceptions of Katherine Parker. Rather, the goal was to further focus participants on the difficulties vs. resilience that stigmatized targets experience to lead participants to construe a second set of scenarios featuring Tyrone Williams in terms of either difficulties or resilience. Moreover, we wanted to include a perspective-taking activity in which the target was still a member of a negatively stereotyped group, but not the same negatively stereotyped group featured in the target scenarios that participants would be asked to complete, to avoid redundancy. Participants were asked to take Katherine’s perspective and write about how she might feel in five different scenarios. In the difficulties condition, the exercise focused on the difficulties Katherine faces as a result of stereotype threat, whereas in the resilience condition, the exercise focused on her resilience in the face of stereotype threat. For instance, one scenario in the difficulties condition was “Meets with her study group to prepare for an exam, and how she feels extra pressure to prove to the group that negative stereotypes about women’s abilities in engineering aren’t true.” The analogous scenario in the resilience condition was “Meets with her study group to prepare for an exam, and how she has learned to perform her best despite the extra pressure to prove to the group that negative stereotypes about women’s abilities in engineering aren’t true.” The goal of these activities was to manipulate participants’ construals of people’s experiences of stereotype threat such that they would evaluate the target scenarios (described below) in terms of the target’s difficulties or their resilience, even though the content of the scenarios was identical across the two conditions.After completing the focus manipulation described above, participants responded to the target perspective-taking exercise, featuring the same Black student used in Study 1. Similar to the previous studies, participants were asked to take the perspective of Tyrone Williams and imagine how he might feel in five different scenarios.did not vary by condition. The scenarios were designed to be ambiguous enough that they could be interpreted either as a sign of the difficulties the target faces or as a sign of their resilience. For instance, one scenario was “Meets with a few other students to work on a group project for his biology class, and wonders what these students think about his abilities.” We expected that participants assigned to the difficulties-focus condition would construe the scenarios in terms of the difficulties Tyrone faces, whereas those assigned to the resilience condition would construe the scenarios in terms of Tyrone’s resilience. Thus, in this way, we manipulated focus on difficulties vs. resilience, while holding all of the information that participants received about the target constant.Importantly, unlike in the previous studies, the content of the target exercise As in Study 1, the dependent measures consisted of participants’ ratings of Tyrone Williams in terms of helplessness (α = .71) and competence (α = .87).M = 2.27, SD = 0.67) viewed the target as more helpless than did those in the resilience condition , t(382) = 2.46, p = .015, d = 0.24.Consistent with our predictions, participants in the difficulties condition  than did those in the resilience condition , t(382) = 3.38, p = .001, d = 0.34.In addition, participants in the difficulties condition viewed the target as less competent  × 3 (difficulties vs. resilience vs. control) between-subjects design. We then examined the extent to which they viewed the target as helpless and competent.Mage = 37.44 years, SD = 11.90) participated in the study in exchange for payment.12 We conducted a series of power sensitivity analyses using G*Power  × 2 (empathy condition: empathy vs. no empathy) between-subjects design. The cover story, procedure, materials, and measures were identical to Study 2, except for the addition of an empathy manipulation.To manipulate empathy for the target, we followed procedures used in previous research . Specifiempathize with Tyrone? ” and “How much do you think you were able to remain objective and detached from Tyrone’s feelings?”). These items were rated on a five-point scale . We formed a composite by reversing the second item and averaging across the two items, with higher scores indicating more empathy.To assess the effectiveness of the empathy manipulation, we included two manipulation check items , and competence (α = .86) with same measures used in Studies 1–3.M = 3.40, SD = 0.68) reported more empathy for Tyrone than participants in the no-empathy condition , F = 133.80, p < .001.As expected for the manipulation check, participants in the empathy condition  = 60.83, p < .001, and no main effect of empathy condition, F = 3.31, p = .129. There was also a marginally significant interaction between focus condition and empathy condition, F = 2.80, p = .061. Given that the interaction was marginally significant, we first report effects for the main effect of focus condition, and then for thoroughness, we report the simple comparisons for the marginally significant interaction.There was a main effect of focus condition on helplessness, M = 2.45, SD = 0.60) than participants in the control condition , and participants in the resilience condition . Participants in the resilience condition also saw the target as marginally more helpless than did those in the control condition, p = .056, d = 0.20.Tukey’s post hoc comparisons for the main effect of focus condition revealed that participants in the difficulties condition viewed the target as more helpless  = 43.83, p < .001, and the empathy condition, F = 19.25, p < .001. Within each empathy condition, the predicted pattern emerged. In the no-empathy condition, participants in the difficulties condition viewed the target as more helpless  than controls, , and participants in the resilience condition . The control and resilience conditions did not differ, p = .184, d = 0.21. Similarly, in the empathy condition, participants in the difficulties condition viewed the target as more helpless  than controls, , and participants in the resilience condition . The control and resilience conditions did not differ, p = .323, d = 0.17. This pattern suggests that the trend towards an empathy by focus condition interaction was driven by the difference between the focus conditions being slightly stronger in the no-empathy condition.Breaking down the marginally significant interaction revealed that the effect of focus condition was significant in both the no-empathy condition, F = 25.87, p < .001, no main effect of empathy, F = 1.09, p = .30 and no empathy by focus condition interaction interaction, F = 0.14, p = .87.There was a main effect of focus condition, M = 3.56, SD = 0.66) perceived the target as less competent than participants in the control condition , and the resilience condition . In contrast, participants in the resilience condition perceived the target as more competent than did participants in the control condition, p < .001, d = 0.34.Participants in the difficulties condition .Six hundred and five White American Mechanical Turk Workers were recruited to participate in the study via TurkPrime . We exclThe procedures of this study were similar to those of the previous studies. Participants were first asked to complete a modified version of the focus manipulation, in which they were randomly assigned to a difficulties condition, a resilience condition, or a control condition. They were then asked to complete the dependent measures.As in Studies 1–4, participants were asked to read about the experiences of Tyrone Williams, a Black college student. Unlike the previous studies, however, the manipulation did not include a stereotype threat article or a perspective-taking exercise. Rather, they were simply asked to “Read about the life about Tyrone Williams.” They were then presented with the same five scenarios used in Studies 1, 3 and 4 describing Tyrone’s experiences. In the difficulties condition, the scenarios described difficulties Tyrone faces as a result of stigma. In the resilience condition, the scenarios described his resilience in the face of difficulties. In the control condition, the scenarios described neutral experiences. In contrast to the previous studies, participants were not asked to write a response to the scenarios; they were simply asked to read them.Perceptions of the target in terms of helplessness (α = .76) and competence (α = .87) were assessed with the same measures used in Studies 1–4.Our measure of admiration was adapted from previous research . ParticiTo assess interest in affiliating with the target in domains in which competence is relevant, we asked participants how willing they would be to engage in different activities with Tyrone in work and academic settings. Specifically, participants responded to the following five items: “How interested would you be in working on an intensive, three-month project with Tyrone?”; “How interested would you be in joining a work group with Tyrone?”; “How interested would you be in asking Tyrone to provide feedback on a report you wrote?”; “How interested would you be in having Tyrone as a partner on a work project?”; “How likely would you be to ask Tyrone for help on a task or assignment if you were struggling?” Responses were given on a five-point scale . A composite was formed by averaging across the five items (α = .95).Consistent with the previous studies, for each of the dependent measures, we conducted a one-way ANOVA and then a series of planned contrasts to examine the effects of the two focus conditions on each of the dependent measures. We predicted that participants in the difficulties-focus condition would view the target as more helpless and less competent and also show less admiration and less interest in affiliating with the target, relative to those in the control condition and those in the resilience condition. In contrast, we predicted that participants in the resilience-focus condition would view the target as more competent and report more admiration and more interest in affiliating with the target, relative to those in the control condition.F = 94.92, p < .001. Participants in the difficulties-focus condition  perceived the target as more helpless than did those in the resilience condition  and the control condition . Participants in the resilience condition also rated the target as more helpless than did those in the control condition , which is consistent with the results of Study 4.Consistent with our predictions, focus condition affected ratings of helplessness, F = 21.47, p < .001. Participants in the difficulties condition  rated the target as less competent than did those in the resilience condition , and those in the control condition . In contrast, participants in the resilience condition rated the target as more competent than did those in the control condition, t(485) = 3.23, p = .001, d = 0.395.Focus condition also affected ratings of the target’s competence, F = 5.25, p = .006. Participants in the difficulties condition  felt less admiration for the target relative to those in the resilience condition . Participants in the difficulties condition did not significantly differ from those in the control condition , however. In contrast, participants in the resilience condition reported greater feelings of admiration for the target than did those in the control condition, t[485] = 2.18, p = .03, d = 0.26.In addition, focus condition affected feelings of admiration for the target, F = 6.34, p = .002. Participants in the difficulties condition  were less interested in affiliating with the target than were participants in the resilience condition . As above, participants in the difficulties condition did not significantly differ from those in the control condition , however. Participants in the resilience condition showed greater interest in affiliating with the target than did those in the control condition, t(485) = 2.94, p = .003, d = 0.35.Finally, focus condition affected interest in affiliating with the target in competence-relevant domains, Study 5 provides additional evidence that focusing on a stigmatized target’s difficulties vs. resilience can have different consequences for perceptions of the target. Indeed, consistent with Studies 1–4, we found that participants who focused on the target’s difficulties perceived him as more helpless and less competent, whereas those who focused on the target’s resilience perceived him as more competent. Moreover, the focus manipulation used in Study 5 did not include a stereotype threat article or an empathy task, suggesting that these components are not necessary for the effects of focusing on difficulties vs. resilience to emerge.In addition, Study 5 suggests that the two ways of approaching stigmatization may affect not only perceptions of helplessness and competence, but also feelings of admiration for the target and interest in affiliating with the target in domains in which competence is relevant. Specifically, participants who focused on the target’s difficulties reported less admiration for the target and less interest in affiliating with the target relative to those who focused on the target’s resilience. In contrast, participants who focused on the target’s resilience showed relatively more admiration for the target and more interest in affiliating with him.Participants in the difficulties condition did not report less admiration and interest in affiliation relative to those in the control condition. It is possible that this pattern emerged because the control condition was less descriptive and contained less information about the target relative to the two experimental conditions. As such, although participants in the control condition may not have seen reason to doubt the target’s competence, they may also not have had strong reasons to admire the target or to want to affiliate with him in competence-relevant domains. Surprisingly, participants in the resilience-focus condition viewed the target as more helpless than participants in the control condition, a finding that did not emerge in the other studies except with marginal significance in Study 4. One possible explanation is that, without the stereotype threat information from Studies 1–4, peoples’ stereotypical perceptions about the helplessness of stigmatized individuals emerge from even a resilience-focused discussion of stigma.Nevertheless, Study 5 provides additional evidence of the different consequences of focusing on difficulties vs. resilience, and suggests that these effects do not emerge only in the context of reading about stereotype threat or completing an empathy task.Conventional wisdom suggests that increasing non-stigmatized or majority group individuals’ understanding of the experiences of members of stigmatized groups will lead to increased positive perceptions of stigmatized individuals and, therefore, more positive intergroup relations. Yet the present research suggests that when such efforts focus narrowly on stigmatized individuals’ experiences of struggle and hardship, they may backfire by leading to more paternalistic perceptions of targets. Indeed, the five studies reported here suggest that focusing only on the difficulties that stigmatized individuals face may lead non-stigmatized individuals to take a paternalistic view of stigmatized targets. The present research also suggests that focusing more broadly on targets’ resilience in the face of their difficulties can improve non-stigmatized individuals’ perceptions of stigmatized targets.construals of that information. Once again, participants in the difficulties condition perceived the target as more helpless and less competent than did those in the resilience condition. In Study 4, we replicated and extended these findings by examining whether it was necessary to instill empathy for the effects to emerge. In this study, we found that, regardless of whether they were asked to empathize with the target or take an objective perspective, Whites who focused on the Black target’s difficulties viewed him as more helpless and less competent than those in a control condition, whereas those who focused on the target’s resilience viewed him as more competent than controls. Thus, although we used interventions to create empathy for targets of stereotype threat as the primary operationalization of the two ways of approaching stigma, Study 4 suggests that the consequences of these two approaches extend beyond the context of empathy interventions. Finally, in Study 5, we examined whether simply reading about a stigmatized target’s difficulties or resilience, without also reading about stereotype threat, would have similar effects to the focus manipulations used in Studies 1–4. Additionally, we measured new outcomes – ratings of affiliation and admiration. This study replicated the findings of Studies 1–4 and found that participants who focused on difficulties still saw the target as more helpless and less competent than controls. Participants who focused on resilience saw the target as more competent and reported more admiration and interest in affiliation relative to controls. Taken together, these four studies suggest that the two ways of approaching stigma – the difficulties-focused approach vs. the resilience-focused approach – have different consequences for people’s perceptions of stigmatized individuals; whereas the resilience-focused approach leads to respectful views of stigmatized targets, the difficulties-focused approach leads to paternalistic views of targets.In Studies 1 and 2, we examined the consequences of an empathy intervention in which White participants were asked to take the perspective of a Black student while focusing on his difficulties or on his resilience in the face of his difficulties (vs. a control condition). Whites who focused on the student’s difficulties subsequently viewed him as more helpless and less competent relative to those in the control condition and the resilience condition. In contrast, Whites who focused on the student’s resilience in the face of his difficulties perceived him as more competent than those in the control condition. In Study 3, we ruled out differences in the ways in which the target was described across conditions as an alternative explanation for the findings. To do so, we held constant the information participants received about the target and manipulated their liked during interracial interactions, racial minorities seek to be respected  were very similar to the items used in the main dependent measures , which may have created demand effects. We do not think that the findings of Studies 1 and 2 can be explained entirely by demand effects, however. Indeed, we removed any wording that was similar to the dependent measures in Studies 4 and 5 and successfully replicated the findings of Studies 1 and 2. Moreover, in Study 3, we gave participants identical information about the targets across the two conditions and found consistent effects.never focus on the difficulties faced by members of stigmatized groups. Neither do we suggest that focusing on their resilience is a panacea for improving intergroup relations. Rather, we highlight one underappreciated negative consequence of efforts to educate non-stigmatized individuals about stigmatization and group-based disadvantage and suggest that such efforts should avoid taking a narrow focus on stigma in which members of stigmatized groups are defined by their struggles and hardships. Instead, discussions of stigma should attempt to foster a broader understanding of stigmatized individuals in which their full humanity is recognized and appreciated.Finally, it is important to note that we are not attempting to characterize the two approaches to stigma as wholly positive or negative. Indeed, there may be positive effects of focusing on a stigmatized target’s difficulties that we have not examined in the present research, such as increasing people’s empathic concern for stigmatized individuals or increasing pro-social behaviors towards stigmatized groups . SimilarThe present research has important implications for theories of intergroup perception. To our knowledge, this work is the first to examine how learning about the difficulties faced by members of stigmatized groups affects people’s perceptions of stigmatized individuals. Although one might think that learning about the difficulties stigmatized individuals face would lead to entirely positive perceptions of these individuals, our research suggests that doing so can lead people to form paternalistic impressions of stigmatized individuals. Ultimately, the difficulties-focused approach may lead to worse outcomes for stigmatized groups insofar as it causes non-stigmatized individuals to behave in more disrespectful ways towards stigmatized individuals or to withhold opportunities and resources from them. In this way, the difficulties-focused approach could ultimately reinforce stigmatization over time. In contrast, focusing on these individual’s resilience in the face of their difficulties may offer a way for non-stigmatized individuals to learn about the experiences of stigmatized groups, and even increase their respect for them, without incurring the negative costs of the difficulties-focused approach.This work also has implications for research on empathy in intergroup contexts. Although a growing body of research has shown that empathy can have negative effects in intergroup contexts , 2012, bdependency-oriented help to members of lower-status groups can act as a status-maintaining mechanism because it places the help-recipients in a low-status, dependent position relative to the help-provider. In contrast, autonomy-oriented helping focuses on empowering individuals to take an active role in addressing their struggles, and is therefore less likely to undermine the recipient’s social status. It is possible that the two approaches to stigmatization examined in the present research may motivate different forms of helping; whereas the difficulties-focused approach may motivate dependency-oriented helping, the resilience-focused approach may motivate autonomy-oriented helping. Future work should explore the connections between these two lines of research.The current work may also be relevant to the literature on dependency vs. autonomy-oriented helping (see Finally, this research has important practical implications. Reducing stigmatization and improving people’s perceptions of stigmatized groups has become an increasingly important issue in society, and there has been a push for programs to increase diversity and reduce the negative effects of stigma across many different types of organizations. It would not be surprising if at least some of these programs have incorporated efforts to educate non-stigmatized individuals about the experiences of members of stigmatized groups. For instance, in one well-known example, Starbucks closed all of its stores for a one-day diversity and inclusion training. As part of this training, Starbucks employees watched a short video entitled “The Story of Access,” that focused on primarily on the struggles and difficulties that Black individuals face, with no discussion of Black people’s broader experiences including their resilience in the face of difficulty. Although these types of programs may be well-intentioned, the present work suggests they may ultimately lead people to form more negative perceptions of stigmatized individuals because they focus exclusively on the difficulties that many members of stigmatized groups face. With a relatively subtle shift, however, such efforts could produce their intended effects. Specifically, organizations should be mindful of the ways they describe the experiences of stigmatized individuals and avoid characterizing them as in need of help or lacking resilience. Instead, organizations should explicitly acknowledge the resilience of stigmatized individuals; doing so may create more respectful environments for stigmatized groups and allow them to thrive.In a society in which group-based stigmatization is an unfortunate reality, the question of how best to foster understanding across group lines is a critical concern. Conventional wisdom suggests that one way to do so is to educate non-stigmatized individuals about the barriers and disadvantages faced by members of stigmatized groups. Yet our research suggests that such efforts must be made with caution. To the extent that these efforts focus narrowly on stigmatized individuals’ experiences of struggle and hardship, they may hinder rather than enhance understanding, and ultimately widen rather than narrow existing group divides.Click here for additional data file.Supplemental material, sj-pdf-1-gpi-10.1177_13684302211009590 for When seeing stigma creates paternalism: Learning about disadvantage leads to perceptions of incompetence by Stephanie L. Reeves, Crystal Tse, Christine Logel and Steven J. Spencer in Group Processes & Intergroup Relations


Angptl7 knockout mice have lower (~2 mmHg) basal IOP compared to wild-type, with a trend towards lower IOP also in heterozygotes. Conversely, increasing murine Angptl7 levels via injection into mouse eyes increases the IOP. We also show that acute Angptl7 silencing in adult mice lowers the IOP (~2–4 mmHg), reproducing the observations in knockout mice. Collectively, our data suggest that ANGPTL7 is important for IOP homeostasis and is amenable to therapeutic modulation to help maintain a healthy IOP that can prevent onset or slow the progression of glaucoma.Glaucoma is a leading cause of blindness. Current glaucoma medications work by lowering intraocular pressure (IOP), a risk factor for glaucoma, but most treatments do not directly target the pathological changes leading to increased IOP, which can manifest as medication resistance as disease progresses. To identify physiological modulators of IOP, we performed genome- and exome-wide association analysis in >129,000 individuals with IOP measurements and extended these findings to an analysis of glaucoma risk. We report the identification and functional characterization of rare coding variants (including loss-of-function variants) in ANGPTL7 associated with reduction in IOP and glaucoma protection. We validated the human genetics findings in mice by establishing that  ANGPTL7 as a promising therapeutic target for intraocular pressure reduction, and protection from glaucoma.Rare variant association data from human genetics combined with in vitro and in vivo functional validation highlight  Classified as a neurodegenerative disease, glaucoma is characterized by the progressive loss of retinal ganglion cells in the eye and thinning of the neuroretinal rim of the optic nerve head. Affected individuals present with visual field loss that is accompanied by increased intraocular pressure (IOP) in the majority of cases2. Primary open angle glaucoma (POAG) is the most common glaucoma subtype and has highest prevalence in individuals of African ancestry .Glaucoma is a leading cause of irreversible blindness, with a global prevalence of 3.54% in individuals 40–80 years of age, and is projected to affect more than 111.8 million people by 20406. Measurable ocular anatomical and physiological features, including low central corneal thickness (CCT), increased cup-to-disc ratio and high intraocular pressure (IOP)7 correlate with increased risk for glaucoma and, like glaucoma8, are highly heritable13. Thus, these quantitative risk factors, when measured on large numbers of individuals, can provide a well-powered dataset for genetic studies to elucidate the etiology of glaucoma risk and progression. The latest genome-wide association study (GWAS) of IOP included more than 130,000 individuals and increased the tally of IOP-associated loci to over 10015. An earlier GWAS16 reported that ~89% of loci associated with IOP at a genome-wide significant level showed directionally consistent effects on glaucoma risk, thus reinforcing the utility of quantitative risk factors in gene discovery for glaucoma.Individuals at greatest risk for POAG are >60 years of age, have a family history of glaucoma, or have high myopia17. Waning efficacy over time and the consequent need for treatment escalation are also observed, perhaps in part because the majority of medications in use do not address pathophysiological changes at the primary site of IOP regulation and aqueous humor egress from the eye, the trabecular meshwork (TM). These limitations result in a large proportion of glaucoma treatment regimens comprising more than one therapeutic agent and patients frequently changing medications or requiring treatment escalation that ultimately involves invasive surgery to maintain a clinically acceptable IOP18. Therefore, a substantial unmet need remains in the treatment of glaucoma to identify new therapeutic targets offering novel mechanisms of action, as well as treatment platforms that may offer increased durability and tolerability without compromising safety.Lowering IOP is the mainstay of all glaucoma therapeutics as IOP continues to be the only modifiable risk factor for the onset or progression of glaucoma. While many effective topical IOP-lowering agents across multiple drug classes are available, they have important drawbacks, including poor compliance due to frequent dosing requirements and side-effectsANGPTL7 as a candidate, consistent with findings reported recently by another group19. In this report, we (1) strengthen the genetic link to glaucoma protection by (i) showing a consistent protective effect of the Gln175His variant in ANGPTL7 across eight cohorts, (ii) identifying a burden of ultra-rare missense variants in ANGPTL7 associated with reduced IOP levels, and (iii) identifying an additional rare ANGPTL7 loss-of-function variant in African-ancestry individuals; we also present (2) in vitro characterization of ANGPTL7 variants identified from genetic analyses; and (3) in vivo results showing that mice lacking Angptl7 have reduced basal IOP, and that even a partial knockdown of Angptl7 with small interfering RNA (siRNA) can lead to lowering of IOP in mice. Our results establish an important role for ANGPTL7 as a physiological regulator of IOP and suggest that it is also amenable to modification by pharmacological tools, making it a compelling target for a glaucoma therapeutic.We performed genetic association analyses of IOP and glaucoma across eight cohorts to identify rare and coding variants that modulate the risk for glaucoma through IOP. This led to the identification of P < 5 × 10−8) of variants in angiopoietin-like 7 (ANGPTL7) with reduced IOP  and the Geisinger DiscovEHR (GHS), including 129,207 individuals of European descent after exclusion of cases with a glaucoma diagnosis  and 26.5% (4.1 mmHg) reduction in median IOP in UKB, respectively  compared to Europeans (MAF = 0.0013%). We performed an association of Trp188* with IOP in African ancestry individuals from UKB and the Primary Open Angle African American Glaucoma Genetics (POAAGG) study, followed by meta-analysis. Trp188* showed a trend towards reduced IOP, similar to Arg177* and Gln175His, but this was not statistically significant . A cross-ancestry meta-analysis of Arg177* and Trp188* variants showed a nominally significant association with reduced IOP , the Malmö Diet and Cancer Study from Malmö, Sweden (MALMO), the FinnGen cohort from Finland, the Estonia Biobank at the University of Tartu, Estonia (EstBB), the HUNT study from Nord-Trøndelag, Norway (HUNT), and the Copenhagen General Population Study/Copenhagen City Heart Study from Copenhagen, Denmark (CGPS-CCHS). A meta-analysis across these eight cohorts showed a significant reduction in glaucoma risk for Gln175His carriers  = 0.77, P = 2.7 × 10−6, Fig. allelic = 0.87, P = 4.1 × 10–1, Fig. ANGPTL7 with reduced IOP and the association of the missense variant with reduced glaucoma risk suggest the hypothesis that loss of ANGPTL7 confers protection against glaucoma, and that this effect is mediated through the regulation of IOP.To understand if carriers of variants in ANGPTL7. We tested the ANGPTL7 variant aggregate for association with 14,050 and 10,032 binary and quantitative traits in UKB and GHS, respectively. No associations reached phenome-wide significance  in GHS. The only significant associations in UKB were with ocular traits  to understand whether other traits were associated with a burden of pLOF and deleterious missense variants in ANGPTL7. We observed a significant association with increased mCRP , corneal astigmatism, and refractive astigmatism (Supplementary Methods) and checked for association with P = 0.08; Supplementary Fig. P < 1 × 10–4; Fig. P < 0.05; Fig. To understand the impact of Gln175His, Arg177*, and Trp188* variants on the expression and secretion of ANGPTL7, we transiently transfected constructs expressing the human wild type (WT), Gln175His, Arg177*, and Trp188* proteins in HEK293 cells. We measured mRNA levels by Taqman, which showed similar Gln175His and Arg177* transcript levels and a trend towards decreased levels of the Trp188* transcript compared to WT . High ANGPTL7 expression was observed in cornea, TM, and sclera in human and African green monkey eyes  deposition and reorganization22 and that ANGPTL7 is increased in aqueous humor of glaucoma patients22, however, the role of ANGPTL7 in IOP regulation is not clear. To investigate this, we injected mAngptl7 protein in mice via intravitreal and intracameral routes and measured IOP over time. Intravitreal injection of murine Angptl7 (mAngptl7) in mice led to an initial drop in IOP followed by, starting on day 4, an elevation in IOP of 4–5 mmHg, a 22–25% increase compared to baseline, that lasted until the end of the experiment on day 7  mice. We confirmed via RNAscope that Angptl7 mRNA was not expressed in any ocular tissue in KO mice whereas it was expressed in TM, cornea, and sclera of WT mice  and WT mice and observed a dose-dependent decrease in IOP across the three genotypes  compared to WT (17.36 ± 0.23 mmHg). Het mice (16.26 ± 0.43 mmHg) showed a smaller  but significant reduction in IOP compared to WT.We generated and characterized ice Fig. . We alsopes Fig. . The meaAngptl7 is highly expressed in cornea and TM and since we used a Tonolab rebound tonometer to measure IOP, we wanted to further confirm that Angptl7 function in the cornea was not affecting IOP. Therefore, we measured the conventional outflow facility using constant flow infusion to understand the relationship between IOP and outflow resistance in Angptl7 KO (n = 7) and WT mice (n = 4). The conventional outflow facility was increased by 21% in KO (25.18 nL/minute/mmHg) compared to WT mice  compared to the PBS and Naïve (no injection) groups  adding six cohorts to the genetic association analyses in UK Biobank and FinnGen that were used in Tanigawa et al, and showing a consistent protective trend in the majority of these for the rare missense variant, Gln175His (rs28991009); (2) identifying an African ancestry-enriched pLOF variant, Trp188*, in ANGPTL7 in addition to Arg177* in Europeans and showing that both variants trend towards protection from glaucoma (3) identifying through exome sequencing and gene burden analyses multiple predicted-deleterious ANGPTL7 variants associated with reduced IOP, indicating that there may be other ultra-rare ANGPTL7 variants that confer protection from glaucoma. Together, these new analyses provide further support for the role of ANGPTL7 in IOP regulation and glaucoma pathogenesis. We also show through cell-based expression assays, that Gln175His, Arg177*, and Trp188* were severely defective in secretion when compared to wild-type and, while not proof, this observation is consistent with the hypothesis that the three variants result in a loss of protein function.In this study, we present genetic and functional evidence for a role for ANGPTL7 in the physiological control of IOP and as a potential target for glaucoma therapy. A recent publication by Tanigawa et al.ANGPTL7 is a secreted glycoprotein that is thought to form homo-oligomers (tri- or tetramers), like other members of the ANGPTL family24. The ANGPTL7 mRNA was first isolated from human postmortem eyes where it shows the highest expression relative to other tissues23. Within the eye, our in situ hybridization results show that ANGPTL7 is expressed most strongly in the cornea and TM, consistent with previous findings22. We have also shown using single-cell RNAseq that ANGPTL7 expression is particularly enriched in the juxtacanalicular tissue (JCT), a region of the TM most important for IOP regulation and generation of aqueous humor outflow resistance26. In addition to being expressed in tissues directly relevant in glaucoma, several lines of evidence have implicated ANGPTL7 in glaucoma pathophysiology. First, elevated levels of ANGPTL7 mRNA and protein were observed in eye tissues from glaucoma patients compared to controls, and under conditions of increased IOP simulated by perfusion of eye anterior segment explants28. Second, ANGPTL7 is one of the most upregulated genes in response to corticosteroid treatment30, which can cause increased IOP in ~40% of general population and ~90% of individuals with POAG32. Third, ANGPTL7 levels are also increased in response to TGF-beta, a growth factor that is thought to modulate the ECM and lead to increased IOP33. Fourth, ANGPTL7 itself can modulate the expression of components of the TM ECM22.Angtpl7 KO mice. Our findings show that increasing mAngptl7 results in increased (~2–4 mmHg) IOP and decreasing mAngptl7 through KO mice reduces basal IOP levels (~2 mmHg). We also observed a trend towards increased conventional outflow facility (~21%) in KO compared to WT mice that was consistent with the decrease in IOP in KO mice. Together, these findings suggest that ANGPTL7 functions in vivo to maintain IOP homeostasis. We further recapitulated the reduction in IOP observed in KO mice by injecting WT mouse eyes with siRNA against Angptl7, which not only replicates the observation in genetic mutant mice but also illustrates that the effect of mAngptl7 on IOP continues post-development and is amenable to modulation by therapeutics in adulthood. Our results of lower IOP in Angptl7 KO mice are highly consistent with observations from human genetics. Based on this, we could extend the siRNA knockdown findings to humans and surmise that inhibition of ANGPTL7 in adulthood could be an efficacious strategy to lower IOP and, eventually, the risk for glaucoma.The above studies suggest a strong correlation of ANGPTL7 expression with glaucoma disease-state, however, they are not evidence for a causal relationship between ANGPTL7 levels and elevated IOP/glaucoma. Human genetics finding of an association between ANGPTL7 loss and lower IOP suggests that ANGPTL7 is physiologically important for IOP regulation. We validated this hypothesis in mice where we performed reciprocal experiments: measuring IOP after increasing ANGPTL7 levels via injection of mAngptl7 into mouse eyes, and after removing all mAngptl7 protein by generating ANGPTL7 associated with reduced IOP were also associated with decreased mean corneal resistance factor (CRF), an output of the Ocular Response Analyzer that reflects the elastic property of the cornea34, and increased mean corneal refractive power (mCRP). mCRP, along with axial length and lens power, contributes to the overall refractive state of the eye35, and increased mCRP (i.e. a steeper cornea) may be indicative of possible myopia or astigmatism. However, as we did not observe evidence of association with myopia or astigmatism in PheWAS, we conclude that the changes to mCRP are either insufficient to result in a disease outcome or are compensated by other changes in traits that were not measured, such as axial length36. While an effect of ANGPTL7 on corneal properties is clearly indicated by the associations with mCRP and CRF, we assert that this effect cannot account for the full extent of IOP reduction observed. Instead, we posit an independent contribution of ANGPTL7 to IOP homeostasis based on the following: (1) The persistent association with reduced IOPcc suggests that there is IOP reduction even after controlling for corneal properties; (2) Angptl7 KO mice eyes show reduced basal IOP without evidence of corneal thinning or other corneal abnormalities; and (3) the association of ANGPTL7 variants with glaucoma protection is a result we would not expect if the reduction of IOP was purely due to ANGPTL7’s effect on corneal anatomy37. Therefore, we believe that ANGPTL7 likely has a pleiotropic effect on both IOP and corneal anatomy/biomechanics in humans.Our human genetics findings indicate that the variants in 38. Consequently, as TM dysfunction progresses in a glaucoma patient, treatment intensification naturally follows18. While further studies are needed to investigate the role of ANGPTL7 in pathways contributing to TM dysfunction and IOP elevation, several lines of evidence point to pro-fibrotic actions at the level of the TM, including its corticosteroid and TGF-beta responsiveness, as well as its enriched expression in JCT TM (discussed earlier).Therapeutic knockdown of ANGPTL7 in patients with open angle glaucoma may present a unique opportunity to not only lower IOP through a novel mechanism of action, but also to intervene in the disease process. Among the most widely used IOP-lowering agents, none directly address pathophysiological changes occurring at the level of TM and instead reduce IOP through mechanisms that reduce the aqueous burden on the conventional pathway, either through suppressing aqueous formation or increasing its outflow via the alternative  pathwayANGPTL7 gene in humans lies within intron 28 of MTOR39, therefore variants in ANGPTL7 could also affect MTOR function. While a possibility, it is unlikely that the effect on IOP and glaucoma is due to MTOR, as: (i) the variants are predicted to be protein-altering in ANGPTL7 whereas there is no obvious predicted functional effect on MTOR; (ii) we show data suggesting that the variants have a functional impact on ANGPTL7; and (iii) we show mouse data that establish a role for ANGPTL7 in IOP regulation.As all studies, this study has limitations. First, we did not account for IOP-lowering medication use in the IOP analysis. We did exclude from the IOP analysis individuals with a glaucoma diagnosis, greatly reducing the effect of medication use, however this would not exclude those individuals on IOP-lowering medications without a glaucoma diagnosis. In addition, the IOP measurement in UKB for most participants was only taken once, and therefore can be prone to errors that a median of multiple measurements might buffer against. Both of the above factors would influence the estimation of the effect of rare genetic variants on IOP. Second, since a glaucoma diagnosis can often be made well after the onset of disease, we can expect that a certain percentage of controls have undiagnosed glaucoma, which may affect the effect size estimate. Third, the In summary, our genetic and pharmacological results indicate that ANGPTL7 participates in the normal physiological regulation of IOP in humans and mice. Since excessive amounts of mAngptl7 protein in the eyes of experimental animals cause IOP to elevate to pathological levels, upregulation of ANGPTL7 in humans may be responsible for the elevated IOP that leads to POAG. Therefore, we propose ANGPTL7 as an excellent candidate to explore as a therapeutic target for POAG.All participants provided informed consent, and studies were approved by the individual IRBs at the respective institutions. UK Biobank has approval from the North West Multi-center Research Ethics Committee (MREC), which covers the UK. It also sought the approval in England and Wales from the Patient Information Advisory Group (PIAG) for gaining access to information that would allow it to invite people to participate. The DiscovEHR study was approved by the Institutional Review Board (IRB) at Geisinger. The BioMe Biobank is an ongoing research biorepository approved by the Icahn School of Medicine at Mount Sinai’s IRB. The Ethical Committee at Lund University approved the Malmo Diet and Cancer Study (LU 51–90) and all the participants provided a written informed consent. The CGPS study (H-KF-01-144/01) was approved by the Ethics Committee of the Capital Region and from the Danish Data Protection Agency. Research at Estonian Biobank is regulated by Human Gene Research Act and all participants have signed a broad informed consent. IRB approval for current study was granted by Research Ethics Committee of University of Tartu, approval nr 236/T-23. For the POAAGG study, approval to enroll and to recontact subjects was obtained from the University of Pennsylvania IRB. The Finngen Biobank was evaluated and approved by the Coordinating Ethics Committee of the Helsinki and Uusimaa Hospital District.40. The GHS MyCode study is a health system-based cohort of patients from Central and Eastern Pennsylvania (USA) recruited in 2007–201941. For IOP association tests in African ancestry individuals, we included 4114 individuals from UKB and 3167 individuals from the Primary Open Angle African-American Glaucoma Genetics (POAAGG) study conducted at the University of Pennsylvania Perelman School of Medicine42. We excluded all participants with a glaucoma diagnosis code (ICD-10 H40) or self-reported glaucoma (UKB field IDs: 6148 and 20002) from IOP analyses.Association with IOP was tested on a total of 101,678 individuals and 27,529 individuals of European ancestry from the United Kingdom Biobank (UKB) and the MyCode Community Health Initiative cohort from Geisinger Health System (GHS), respectively. The UKB is a population-based cohort study of people aged between 40 and 69 years recruited through 22 testing centers in the UK between 2006 and 2010ANGPTL7 variants with glaucoma was tested in 8 studies: UKB, GHS, Mt. Sinai BioMe cohort (SINAI), the Malmö Diet and Cancer study (MALMO)43, the Estonia Biobank (EstBB)44, The Trøndelag Heath Study (HUNT)45, FinnGen, a study from Finland, and the Copenhagen General Population Study and the Copenhagen City Heart Study (CGPS-CCHS)46. We had, in total, up to 40,042 cases  and 947,782 controls of European ancestry, and 5153 cases  and 21,650 controls of African ancestry in glaucoma analyses.Association of 47. For this study, we focused on IOPg as this measurement is the most comparable to IOP measurements in other cohorts, and herein IOPg will be referred to as IOP. IOP in POAAGG was measured using a GAT. In GHS, IOP measurements were obtained from several instruments including GAT, Tono-pen and I-Care, which are correlated with IOPg readings from the Ocular Response Analyzer48. For GHS individuals who were not prescribed any IOP medications, we used the median of all IOP measurements available. For individuals who had an IOP medication prescribed, we used the median of IOP measurements available preceding the start date for IOP medications (if available). Individuals for whom we did not have non-medicated IOP values were excluded from the IOP genetic analyses. For association analyses of IOP, we excluded individuals with: (1) a glaucoma diagnosis; (2) IOP measures that were more than 5 standard deviations away from the mean; (3) more than a 10-mmHg difference between both eyes. We derived a mean IOP measure between both eyes for each individual. IOP of only one eye was used in instances where IOP measures for both eyes were not available.IOP in UKB was measured in each eye using the Ocular Response Analyzer . Participants were excluded from this test if they reported having eye surgery in the preceding 4 weeks or having an eye infection. The Ocular Response Analyzer calculates two forms of IOP, a Goldmann-correlated IOP (IOPg) and a corneal-compensated IOP (IOPcc). IOPg most closely approximates the IOP measured by the Goldmann Applanation Tonometer (GAT), which has been the gold standard for measuring IOP, while IOPcc provides a measure of IOP that is adjusted to remove the influence of corneal biomechanics42.Details on glaucoma definition in each cohort are given in the Supplementary Methods. In brief, glaucoma cases in GHS, SINAI, MALMO, HUNT, EstBB, FinnGen (v.R3) and CGPS-CCHS were defined by the presence of an ICD-10 H40 diagnosis code in either outpatient or inpatient electronic health records. In UKB, glaucoma cases were defined as individuals with either an ICD-10 H40 diagnosis or self-reported glaucoma (UKB field ID: 6148 or 20002). In the POAAGG cohort, glaucoma cases and controls were classified based on an ophthalmic examination by glaucoma specialists, and glaucoma suspects were also included in the cases50 as described in Supplementary Methods. We estimated the association with IOP and glaucoma of genetic variants or their gene burden using REGENIE v1.0.4351 , SAIGE52  or logistic regression (CGPS-CCHS). Analyses were adjusted for age, age2, sex, an age-by-sex interaction term, experimental batch-related covariates, and genetic principal components, where appropriate. Cohort-specific statistical analysis details are provided in Supplementary Methods. Results across cohorts were pooled using inverse-variance weighted meta-analysis. Details on the PheWAS analysis conducted in UKB and GHS are provided in Supplementary Methods. Western blotting and ELISA analyses were repeated on three independent biological replicates and data are presented as mean ± SEM. Technical replicates (n = 3) were run for the ELISA analysis. P values were calculated by one-way ANOVA with Tukey’s multiple comparison test for multiple groups analysis  encoding the following proteins: ANGPTL7 WT, Gln175His, Arg177* and Trp188*. After 24 h, the media was changed with 2% FBS OptiMEM. The following day, the cells were collected in RIPA buffer, supplemented with protease and phosphatase inhibitors (BRAND) or TRIzol reagent (Invitrogen) for protein and RNA analysis, respectively. The supernatants were transferred to an Eppendorf tube and immediately flash frozen for downstream protein analysis. Western blot analysis was performed using a rabbit polyclonal antibody against ANGPTL7 at 1:1000 dilution (10396-1-AP ProteinTech), using standard procedures. ANGPTL7 was quantified by ELISA according to manufacturer’s instructions (LS-F50425 Life Sciences). The cell lysates were diluted 1:1000. The supernatants were diluted 1:10,000. The ELISA plate was read at 450 nm via SpectraMax M4 plate reader (Molecular Devices).HEK293 cells, derived within Regeneron, were cultured in DMEM media 4.5 g/L D-Glucose, (+) L-Glutamine, (–) Sodium Phosphate, (–) Sodium Pyruvate supplemented with 10% FBS and 1% Penicillin-Streptomycin-Glutamine (Invitrogen), at 37 °C in a humidified atmosphere under 5% COTotal RNA was extracted using TRIzol reagent (Invitrogen) and RNeasy kit (Qiagen) according to manufacturer’s instructions and treated with RNase-free DNase I (Promega). cDNA was synthesized using Superscript VILO cDNA synthesis kit (Invitrogen). Taqman analysis was performed using TaqMan Fast Advanced Master Mix (Applied Biosystems) in a QuantStudio 6 Flex (Applied Biosystems) and commercially available primers and probes for ANGPTL7 (Hs00221727—Applied Biosystems) and GAPDH (Hs02786624_g1—Applied Biosystems).Angptl7–/– mice, on 63% C57BL/6NTac and 37% 129SvEvTac background, were generated by Regeneron Pharmaceuticals using the VelociMouse® technology53. Heterozygous mice (Angptl7+/−) were bred to generate age-matched wild-type, het and KO littermates that were used for experimentation at 3-4 months of age (mixed gender). Ocular anatomy in these mice was characterized using optical coherence tomography. Detailed methods on generation and characterization of KO mice are provided in Supplementary Methods. For in vivo siRNA experiments, we used C57BL/6 J male mice, 3-4 months old, from Jackson Labs.All animal protocols were approved by the Institutional Animal Care and Use Committee in accordance with the Regeneron’s Institutional Animal Care and Use Committee (IACUC) and the Association for Research in Vision and Ophthalmology (ARVO) Statement for the Use of Animals in Ophthalmic and Vision Research. 56. When testing Angptl7 siRNAs, IOPs were measured in each eye before then start of experiment and then every week until end of study. IOP measurements for both eyes were completed within 3–5 min. Six correct single measurements were done on each eye to generate one IOP reading. We took five IOP readings for each eye and used the average of those readings at each time-point.Mice were anesthetized and IOP was measured in both eyes using a TonoLab rebound tonometer  before the start of Angptl7 injection and every day afterwards for six days58. Mice were anesthetized by using a 100/10 mg/kg ketamine/xylazine cocktail. A quarter to half of this dose was administered for maintenance of anesthesia as necessary. One to two drops of proparacaine HCl (0.5%) (Bausch + Lomb) were applied topically to both eyes for corneal anesthesia. The anterior chambers of both eyes were cannulated by using a 30-gauge needle inserted through the cornea 1–2 mm anteriorly to the limbus and pushed across the anterior chamber to a point in the chamber angle opposite to the point of cannulation, taking care not to touch the iris, anterior lens capsule epithelium, or corneal endothelium. Each cannulating needle was connected to a previously calibrated  flow-through BLPR-2 pressure transducer  for continuous determination of pressure within the perfusion system. A drop of genteal (Alcon) was also administered to each eye to prevent corneal drying. The opposing ends of the pressure transducer were connected via further tubing to a 1 ml syringe loaded into a microdialysis infusion pump . The tubing, transducer, and syringe were all filled with sterile DPBS (Gibco). Signals from each pressure transducer were passed via a TBM4M Bridge Amplifier (WPI) and a Lab-Trax Analog-to-Digital Converter (WPI) to a computer for display on a virtual chart recorder . Eyes were initially infused at a flow rate of 0.1 μl/min. When pressures stabilized within 10–30 min, pressure measurements were recorded over a 5-min period, and then flow rates were increased sequentially to 0.2, 0.3, 0.4, and 0.5 μl/min. Three stabilized pressures at 3-minute intervals at each flow rate were recorded. C in each eye of each animal was calculated as the reciprocal of the slope of a plot of mean stabilized pressure as ordinate against flow rate as abscissa.Aqueous humor outflow facility (C) was measured by using our constant flow infusion technique in live miceA 33-gauge needle with a glass microsyringe  was used for injections of Angptl7 protein/siRNA into mice eyes. For intravitreal injections, the eye was proptosed, and the needle was inserted through the equatorial sclera and into the vitreous chamber at an angle of approximately 45 degrees, taking care to avoid touching the posterior part of the lens or the retina. Angptl7 protein  or siRNA  or PBS (1uL) was injected into the vitreous over the course of 1 minute. The needle was then left in place for a further 45 s (to facilitate mixing), before being rapidly withdrawn. siRNA sequences for all six probes tested are provided in Table Further information on research design is available in the Peer Review FileSupplementary InformationDescription of Additional Supplementary FilesSupplementary Data 1Supplementary Data 2Reporting Summary


Osteoporosis is a progressive bone disorder with a higher incidence in the elderly and has become a major public health concern all over the world. Therefore, it is urgent to investigate the mechanisms underlying the pathogenesis of osteoporosis. In this study, the osteoporosis animal model was established, and then rat bone marrow mesenchymal stem cells (rBMSCs) were cultured. The results showed that PHF8 expression was decreased in osteoporosis rats compared to controls. Overexpression of PHF8 promoted BMSC osteogenic differentiation and the expression of osteogenesis-related genes. In addition, the Wnt/β-catenin signaling pathway in BMSCs was inhibited in osteoporosis rats, which was rescued by overexpression of PHF8. After treatment with the Wnt pathway antagonist, the improved osteogenic differentiation of BMSCs induced by overexpression of PHF8 was blocked. Collectively, our data revealed that the decreased expression of PHF8 in osteoporosis rats suppressed the osteogenic differentiation of BMSCs, which was then restored by PHF8 overexpression. Furthermore, the inhibition of the Wnt/β-catenin signaling pathway in BMSCs suppressed osteogenic differentiation. Thus, these findings indicated that PHF8 plays a role in osteogenic differentiation through the Wnt/β-catenin signaling pathway.  The epiBone marrow mesenchymal stem cells (BMSCs) are considered to be multipotential progenitor cells with strong proliferation ability that are able to differentiate into various types of cells, including osteoblasts, chondrocytes, adipocytes, osteocytes, and myoblasts . BMSCs hIn-silico studies provide insight into the effect of each compound and reveal which perturbations may deregulate the basal healthy state of cells and tissues [Activation of Wnt/β-catenin signaling pathways can regulate cell proliferation and survival . When th tissues ,22. Stru tissues , which p tissues ,25.in vitro cell experiments. Using an animal model of osteoporosis and in vitro BMSCs culture, we examined the expression of PHF8 and the osteogenesis efficiency of BMSC. We found that PHF8 regulated osteogenic differentiation in rat bone marrow mesenchymal stem cells (rBMSCs) via the Wnt/β-catenin signaling pathway. Then this study identified a new target, PHF8, regulating BMSC osteogenic differentiation, providing crucial insight into the mechanism underlying the pathogenesis of osteoporosis and new therapeutic options for osteoporosis.In this study, we aimed to explore the role of PHF8 in osteoporosis. Through ovariectomy, we constructed an animal model of osteoporosis and isolated BMSCs from rat femur and tibial epiphyseal regions for 22.1Female Wistar rats were purchased from Beijing Huafukang Bioscience Co., Inc  and housed at specific pathogen-free (SPF) conditions.Twelve female SPF Wistar rats (aged 7 months and weighing 400–500 g) were randomly divided into sham and ovariectomy (OVX) groups. A bilateral ovariectomy was performed to establish an osteoporosis model for the rats in the OVX group, while a sham surgery in which the ovaries of rats were only exteriorized but not resected was carried out on the sham group as previously described ,27. Six Ethical approval: The research related to animal use has been complied with all the relevant national regulations and institutional policies for the care and use of animals and was approved by the Ethics Committee of the Shanghai Jing’an District Zhabei Central Hospital.2.2Rat BMSCs (rBMSCs) were obtained from the femur and tibial epiphyseal regions of rats as previously described . rBMSCs 2.3The femurs of rats from the sham and OVX groups were fixed with 4% paraformaldehyde, decalcified with 0.5 M ethylenediamine tetraacetic acid, embedded in paraffin blocks, and sectioned at a thickness of 4–5 μm. Then, hematoxylin and eosin (H&E) staining was performed on the sections .For cellular immunofluorescence (IF), the antibody against β-catenin  was used. Cells were stained with 4’,6-diamidino-2-phenylindole reagent for nuclear staining.Osteogenesis was detected by Alizarin Red S  as previously described . BrieflyStaining was observed by microscopic  and photographed. Histological results were analyzed by ImageJ software.2.4The cells and tissue homogenate were lysed in radio-immunoprecipitation assay buffer with protease and phosphatase inhibitors. The concentrations of the supernatant were determined by a bicinchoninic acid (BCA) protein detection kit  . Equal a2.5Total RNA was extracted from cultured cells using Trizol reagent  according to the manufacturer’s instructions . The cDN2.6ALP activity was detected in the cell supernatants using a colorimetric assay kit  . ALP act2.7post hoc test for multiple comparisons and a Student’s t-test for binary comparisons was applied. Otherwise, a Kruskal–Wallis test with Dunn’s post hoc test for multiple comparisons and a Mann–Whitney U-test for binary comparisons were performed. P < 0.05 was considered statistically significant.All data analysis and visualization were carried out using GraphPad Prism software (version 9.0). The data were expressed as the mean ± standard deviation of at least three experimental replicates. A Shapiro–Wilkinson test was used to confirm the normality of the data. If the data were normally distributed, ANOVA with Tukey’s 33.1To confirm the establishment of the osteoporosis rat model, bone microarchitectures in the femur were examined by H&E staining. We found that trabecular bone mass was markedly decreased in the OVX group compared with sham groups . Further3.2To evaluate the function of PHF8, rBMSCs were isolated and transfected with a PHF8 overexpression plasmid vector. After transfection, the increased expression of PHF8 in rBMSCs was confirmed by Western blotting . Of sign3.3To further investigate the effect of PHF8 on rBMSCs, the mRNA and protein expression levels of key factors related to osteogenic differentiation were detected by real-time PCR  and west3.4The Wnt/β-catenin signaling pathway plays a fundamental role in bone remodeling, such as in osteoporosis . Whether3.5To further explore the mechanisms of PHF8 underlying osteogenic differentiation, BMSCs were treated with the Wnt pathway antagonist, DDK1. Alizarin Red S staining results indicated that treatment with DDK1 weakened rBMSC osteogenic differentiation induced by overexpression of PHF8 . Meanwhi4Osteoporosis is a progressive bone disorder with an increasing incidence worldwide, especially in the elderly, and it is characterized by the systemic loss of bone mass and microarchitecture, which increases the risk of fragility fractures ,2,3,35. Histone demethylases have emerged as key regulators of biological processes, and the disorder of histone demethylases is associated with a variety of cancers , neuronaThe Wnt/β-catenin signaling pathway has been found to regulate a series of biological processes . InapproThere are several limitations to this study. The major limitations of this study were the lack of human specimens that could be used to further verify the effect of PHF8 on human BMSCs. Although animal and cell experiments may closely mimic the characteristics of osteoporosis disease, the consistency would be enhanced if further verification is conducted. While this report was the result of preliminary research, we will utilize human specimens in further research.In summary, we first found that decreased PHF8 expression deturbed osteogenic differentiation of BMSCs and inhibited the Wnt/β-catenin signaling pathway in osteoporosis rats. In addition, PHF8 overexpression promoted osteogenic differentiation of BMSCs, which was suppressed by inhibition of the Wnt/β-catenin signaling pathway. Thus, these results uncovered that PHF8 perform its promoting function of osteogenic differentiation via the Wnt/β-catenin signaling pathway. Thereby, this study provides novel insights into the mechanisms underlying the development of osteoporosis, which may provide a potential therapeutic target in the future.


Bearing on artificial hip joint experiences friction, wear, and surface damage that impact on overall performance and leading to failure at a particular time due to continuous contact that endangers the user. Assessing bearing hip joint using clinical study, experimental testing, and mathematical formula approach is challenging because there are some obstacles from each approach. Computational simulation is an effective alternative approach that is affordable, relatively fast, and more accessible than other approaches in examining various complex conditions requiring extensive resources and several different parameters. In particular, different gait cycles affect the sliding distance and distribution of gait loading acting on the joints. Appropriate selection and addition of gait cycles in computation modelling are crucial for accurate and reliable prediction and analysis of bearing performance such as wear a failure of implants. However, a wide spread of gait cycles and loading data are being considered and studied by researchers as reported in literature. The current article describes a comprehensive literature review adopted walking condition that has been carried out to study bearing using computational simulation approach over the past 30 years. Many knowledge gaps related to adoption procedures, simplification, and future research have been identified to obtain bearing analysis results with more realistic computational simulation approach according to physiological human hip joints. Computational simulation; Human hip joint; Hip joint prosthesis; Hip resurfacing; Total hip arthroplasty; Walking condition. This surgery has been considered the best option to relieve damage human hip joint to reduce pain, enhance joint functional, and improve life quality of patients , 2, 3. TAlthough hip joint replacement surgery is considered one of the biggest developments in orthopaedics in the last few decades, this operation has not been entirely successfully studied from a mechanical perspective, so it requires further studies. The service life of hip joint prosthesis is generally limited to range of 15 years that does not provide satisfaction for young patients under 60 years of age with longer life expectancy, more than 40% expect implant life to exceed 20 or 25 years . The sucIn implant's components, the bearing couple that consist of femoral head and acetabular cup play an essential role as load-bearing and provide movement articulation that continuously in contact for every user's activity . BearingIn performing various bearing couple studies on medical implant, four approaches can be used, namely clinical study , experimComputational simulation approach enables to overcome the most common problems found in the first three approaches. This approach carried out by various researchers currently uses finite element method to investigate bearing couple on hip joint arthroplasty with various parameters for further exploration , 32, 33.Over the past 30 years, various efforts to develop bearing couples of artificial hip joints by many researchers using computational simulation presented in this review paper have been studied various aspects, such as geometry , materiaThe objective of present review article is to comprehensively summarizes the adoption of walking conditions in previous studies using computational simulations to assess couple bearing in total hip prosthesis and hip resurfacing. The previous literature over the past 30 years (1992–2022) from Scopus database has been collected and further examined to understand adopted walking conditions that have been done using the computational simulation approach. In-depth information about adoption procedures, simplification, and future research has been presented in this review for filling knowledge gaps in the literature.2Computational simulation based on finite element on bearing of hip joint prosthesis have been attempted to represent how bearing working in actual condition, when used by patient, so this approach can not give significant difference on results from clinical study or experimental testing. Therefore, it is necessary to establish a variety of input data and boundary conditions that are as realistic as possible to achieve results that are closer to actual condition. The use of realistic loading originating from physiological of human hip joint has been carried out in various previous studies by adopting physiological this joint in performing various activities. With rationalization walking is the most common human activity, most realistic loading considered by many researchers to study bearing of hip joint implant is adoption of walking condition , 58, 59.With regards to physiological human hip joint during walking, there are loading and motions that work in three dimensions, shown in In adopting walking condition, three components must be considered for computational simulation approach, there are loading, motions, and cycle described in In general, research flow of computational simulation approach using finite element method on artificial hip joint's bearing under walking conditions is described in 3In computational simulation approach to assess bearing on hip joint prosthesis, two aspects affect the results under walking conditions, there are adopted walking condition and walking condition reference. In first aspect, it is divided into simplified and physiological walking conditions. For the second aspect, it is divided into ISO, published literature, and independent measurement. Therefore, it will not be able to get the same simulation results from one researcher to another, even examining same type of hip joint bearing under walking conditions in computational simulation studies. For more detailed, it is described in Over the past 30 years, many researchers have conducted several efforts to develop bearing of hip joint arthroplasty using the finite element method under walking condition. Several computational simulation software selected to conduct finite element analysis, like ANSYS , ABAQUS The adoption of walking conditions in previous studies has been carried out by considering the loading components, both 3D load , 2D loadFor the adoption of walking conditions on motion components, previous studies have considered several types, ranging from FE-AA-IER , FE 75]75], and Simplification of the cycle for an adopted walking condition has been carried out previously by dividing the walking cycle into 16 , 20 68]68], 25 , 28 47]47], 32 , and 41 The simplification of adopted walking condition is also influenced by the finite element model used to study couple bearings. The use of a three-dimensional finite element model allows researchers to adopt physiological walking conditions without any simplification of the loading, motions, and cycle components, as done by Affatato et al. , Kang etSimplification of adopted walking condition can certainly affect the results of computational simulation. Using walking condition data from Bergmann et al.  shown inDifferences in walking condition reference can also affect the results obtained from computational simulation studies, even though they are both simplified or physiological walking conditions. Liu et al.  studied 4Considering at research on artificial hip joint's bearing over the past 30 years with computational simulation approach, many researchers have used simplified walking condition as described in Walking condition has been adopted by many researchers mostly still using “normal” condition with unspecific subjects , 67, 102Apart from that, in daily human life, most common activities are walking. However, there are still many other activities, such as sit down/get up , load tr5Many researchers have conducted several studies in developing bearing of artificial hip joints, both total hip arthroplasty and hip resurfacing through computational simulation approach using finite element method to avoid various obstacles from clinical study, experimental testing, and mathematical formula. Over the past 30 years, researchers have adopted walking condition to investigate bearing for obtaining results in solid, fluid, or both domains. Unfortunately, to alleviate heavy computational process, various studies have simplified walking conditions, whether on loading, motions, or cycles that can affect the results and lead researchers to misinterpret the results. Adoption of walking conditions of specific subjects needs to be done to develop medical implants for better results so as to minimize implant failures that require revision surgery. Considering other human activities should also be contemplated in conjunction with normal walking to obtain more realistic results.All authors listed have significantly contributed to the development and the writing of this article.The research was supported by World Class Research UNDIP number 118-23/UN7.6.1/PP/2021.No data was used for the research described in the article.The authors declare no conflict of interest.No additional information is available for this paper.