Human T-cell leukemia virus type 1 (HTLV-1) was the first retrovirus described as a causative agent for human disease. In the field of ophthalmology, a close relationship between HTLV-1 infection and uveitis was identified through a series of clinical and laboratory studies in the late 1980s–1990s. Since then, HTLV-1-related ocular manifestations such as keratoconjunctivitis sicca, interstitial keratitis, optic neuritis and adult T-cell leukemia/lymphoma (ATL)-related ocular manifestations have continuously been reported. During the three decades since the association between HTLV-1 and ocular pathologies was discovered, ophthalmic practice and research have advanced with the incorporation of new technologies into the field of ophthalmology. Accordingly, new findings from recent research have provided many insights into HTLV-1-associated ocular diseases. Advanced molecular technologies such as multiplex polymerase chain reaction (PCR)/broad-range PCR using ocular samples have enabled rapid and accurate diagnosis. Advanced ophthalmic technologies such as widefield fundus camera and optical coherence tomography (OCT) have clarified various features of HTLV-1-associated ocular manifestations, and identified characteristics such as the “knob-like ATL cell multiple ocular infiltration” (KAMOI) sign. Advanced drug delivery methods such as intravitreal injection and sub-Tenon injection have led to progress in preventing disease progression. This article describes global topics and the latest research findings for HTLV-1-associated ocular diseases, with reference to a large-scale nationwide survey of ophthalmologists. Current approaches and unmet needs for HTLV-1 infection in ophthalmology are also discussed. In fact, we humans typically obtain 80% of our information on the outside world through sight. Visual impairment thus exerts a major impact on quality of life. To see things clearly, smooth light penetration throughout the eye is essential. Visual recognition can be obtained when light from the outside world passes through the cornea, anterior chamber, lens, and vitreous to reach the retina. Thereafter, photonic stimuli are converted into neural signals that reach the brain through the optic nerve. Disturbance of even a single part of this transmission pathway from the eye results in visual impairment.Human T-cell leukemia virus type 1 (HTLV-1) is a retrovirus that has been attracting worldwide attention since it was found that fully half of Aboriginal adults in central Australia are infected by the virus . LookingIn this article, I discuss: (1) the history of HTLV-1-associated ocular diseases; (2) the global distribution of HTLV-1-related eye diseases; (3) recent advances in ophthalmology and their application to HTLV-1-associated ocular diseases; (4) current knowledge regarding HTLV-1-associated ocular diseases; (5) current concerns; and (6) unmet research needs. I believe this article will prove useful for organizing disparate information on HTLV-1-associated ocular diseases.To date, many ophthalmologists have reported a variety of ocular manifestations related to HTLV-1 infection. From the latter half of the 1980s to the early 1990s, the relationship between HTLV-1 and ocular inflammation, i.e., uveitis, has been reported from the perspective of clinical research among HTLV-1 carriers . ConfirmAs for other ocular manifestations among HTLV-1 carriers, a relationship between HTLV-1 and Sjögren’s syndrome/keratoconjunctivitis sicca (KCS) was reported in the early 1990s . InterstHTLV-1 is known to cause HAM, a progressive neurologic disease. HU was reported to accompany HAM in the late 1980s . SjögrenATL is one of the most aggressive leukemias, and has been identified to be caused by HTLV-1 infection. Ocular manifestations among ATL patients started to be reported around the late 1980s–1990s . HoweverHTLV-1-associated ocular disease is estimated to occur all over the world, corresponding to the geographical distribution of HTLV-1 infection . Here, tThe first evidence of links between HTLV-1 and ocular diseases was reported from Japan. The most frequently reported HTLV-1-associated ocular disease is HU . OphthalAs for other ocular manifestations, the relationship between Sjögren’s syndrome/KCS and HTLV-1 infection was first noted in Japan . CornealHU has been reported to accompany HAM in Japan . SjögrenATL-related ocular diseases are rare , resultiScrutinizing these reports from an ethnic perspective, most HTLV-1 ocular manifestations have been identified in Japanese and Caribbean populations. These ethnic groups are known to show high rates of HU, HU accompanied by HAM, corneal lesions, and ATL-related ocular manifestations. Ophthalmic researchers in Brazil indicated that Caucasian and mixed-ethnicity groups also experience HU and KCS . AfricanThe traditional fundus camera can capture around 30% of the retinal field with full-field flash illumination. A widefield fundus camera, the Optos© , was developed in Scotland and became commercially available in the 2000s. The widefield fundus camera can capture more than 80% of the fundus in a non-invasive manner and without requiring mydriasis. The advantage of the widefield fundus camera is visualizing a retinal peripheral lesion in one picture, as this is not possible with a traditional fundus camera. Such cameras are now an essential tool among ophthalmologists for the identification of peripheral retinal and vascular pathologies. The concept underlying widefield imaging has been applied to other fundus-visualizing technologies, such as widefield fluorescein angiography for retinal vessel analysis, widefield indocyanine green angiography for choroidal vessel analysis, and widefield autofluorescence for retinal pigment epithelium analysis.Imaging of ocular lesions is important to identify HTLV-1-associated ocular lesions. HU results in retinal vasculitis, while ATL induces intraocular infiltration and cytomegalovirus (CMV) retinitis at the periphery of the retina. For instance, careful attention should be directed toward possible necrotic lesions of CMV retinitis arising at the peripheral retina. To follow-up such CMV lesions in the periphery of the retina, widefield fundus images are indispensable .Optical coherence tomography (OCT) is one of the most important modalities applied in ophthalmology. OCT is a non-invasive diagnostic technique that enables visualization of the retina in a cross-sectional view. OCT has greatly evolved the practice of ophthalmology, as cross-sectional mapping of the retina has provided large amounts of information and new insights.OCT is useful for the diagnosis of HTLV-1-associated ocular lesions such as HU and ATL. For example, HU is sometimes accompanied by cystoid macular edema (CME), which significantly affects visual acuity. OCT can clearly reveal the features of CME . As for Marked advances in molecular technologies have been made in the last decade. The diagnostic methods of uveitis in clinical practice have been significantly changed by the application of molecular biological technologies, particularly polymerase chain reaction (PCR).PCR allows accurate analysis of small volumes of ocular samples (such as aqueous humor or vitreous fluid). However, several applications of the technique have been necessary to check for the various causative pathogens. Recently, our group developed multiplex and broad-range PCR methods , 2013. TAs for the diagnosis of HTLV-1-associated ocular lesions, especially among ATL patients, our PCR method is quite useful. ATL-related ocular manifestations are very complicated, because the ocular presentations of infection and infiltration are sometimes difficult to distinguish. Identifying or excluding pathogenic agents is a very important clue for the diagnosis of ATL ocular manifestations. If pathogenic agents can be identified, pathogen-specific drugs  can be given promptly, leading to protection against ocular tissue damage by pathogens. If pathogenic agents fail to be identified, ATL infiltration should be considered. As ocular infiltration of ATL cells is rapid and results in severe destruction of intraocular tissues, addressing this with adequate treatment is critical to prevent vision loss.Delivery of pharmacotherapies by intravitreal injection (IVI) represented a turning point in the treatment of various of ocular conditions, including age-related macular degeneration, diabetic macular edema, proliferative diabetic retinopathy, retinal vein occlusion, pathological myopia, and uveitis. In the early 1900s, IVI was introduced for procedures addressing retinal detachment. In recent years, the use of IVI has grown exponentially, due to the progressive expansion of clinical applications.ATL patients often have multiple organ disorders such as kidney failure due to the disease itself or side effects of drugs, and systemic treatment should be avoided in such cases. In this situation, IVI of the drug offers a useful delivery method to lesions in the eye, while reducing overall burden on the body. Rapid progression of a lesion should stop with immediate treatment and the most effective approach for this situation is delivering the drug through IVI, as pharmaceutical agents are able to reach the lesion directly. Progressive ocular lesions such as ocular infiltration of ATL easily leads to vision loss and thus countermeasures should be enacted immediately . DeliverAs for relatively new delivery methods, sub-Tenon injection has been noted. This delivery method is mainly applied for HU and the accompanying CME. In cases of severe vitreous opacity and CME in HU, in which eye drops or oral steroid administration are not sufficient, injection of steroid such as triamcinolone acetonide into the sub-Tenon space appears much more effective.HTLV-1 uveitis (HU) was the first ocular disease directly associated with HTLV-1 infection from the perspective of clinical and basic research in the early 1990s. According to the anatomical classification of intraocular inflammation, most cases of HU are classified as intermediate uveitis, followed by posterior uveitis. The main clinical feature of HU is the moderate to high infiltration of inflammatory cells into the vitreous, resulting in vitreous opacities . As a reThe peak age at uveitis onset was in the 50s followed by the 60s for women, and in the 60s followed by the 70s for men, according to one recent survey , which sAs for ocular complications secondary to HU, cataracts represent the most common ocular complication, followed by glaucoma, CME, and epiretinal membrane . In termA series of studies has been conducted on the pathogenesis of HU. According to analyses of intraocular fluid, HTLV-1-infected cells infiltrate the eye, where HTLV-1 provirus  and viruAccording to basic research, HTLV-1 has the potential to infect human ocular cells . These iAs for animal models, the first animal model of HU was established in the early 1990s. HTLV-1-infected rabbits showed uveitis after a latency of 3.5 years. The ophthalmic features of rabbits were quite similar to HU and HTLV-1-associated ocular conditions of humans from the perspectives of corneal opacity, anterior inflammation, vitreous opacity, and retinal destruction . Using aBased on the above series of basic studies, the current explanation of the mechanisms underlying HU is as follows. In HU patients, the number of HTLV-1-infected cells increases in the peripheral blood and infected cells significantly accumulate in the eye. Accumulated HTLV-1-infected cells in the eye produce various inflammatory cytokines and chemokines, leading to damage to ocular tissues. From another perspective, HTLV-1-infected RPE cells contribute to the breakdown of immune homeostasis in the eye. Cross-reactivity between HTLV-1 and retinal antigens would also contribute to HU. However, what causes the increase in infected T cells in peripheral blood and why accumulation occurs in the eye both remain unclear. Further investigation is needed to identify the mechanisms of HU in detail.In the clinic, HU is diagnosed by the presence of uveitis, confirmation of HTLV-1 infection , and exclusion of other etiologies of defined uveitis , 2017. FHU is considered to be caused by inflammatory cytokines produced from HTLV-1-infected cells, which significantly accumulate in the eyes of the patient. Topical and/or oral corticosteroid treatment is therefore effective to treat HU patients, suppressing cytokine production by the HTLV-1-infected T cells causing intraocular inflammation , 2017. CAs for ocular complication secondary to HU, advanced cataracts require cataract surgery. Cataract surgery in uveitis must be performed with a low level of aggression and a high level of safety  by selecKeratoconjunctivitis sicca (KCS), also known as dry eye syndrome, involves chronic, bilateral discomfort of the conjunctiva and cornea due to inadequate lubrication and a reduced tear meniscus with a tear breakup time under 10 s. KSC is known to be associated with systemic diseases such as Sjögren’s syndrome, rheumatoid arthritis (RA) and lupus.In the late 1980s, basic research in animal models showed tear film alteration with Sjögren’s syndrome in HTLV-1-related transgenic mice . From thAs for treatment, the definitive treatment for KCS in HTLV-1 carriers has not been established. The current approach is symptomatic therapy. Lubrication of the ocular surface is needed to avoid the development of superficial punctate keratitis and corneal ulcers.Chronic interstitial keratitis (IK) related to HTLV-1 infection was first reported in the early 2000s . KeratitThe mechanism underlying these corneal lesions could be linked to lymphoplasmacytic infiltration of the stroma leading to corneal opacities . CornealOptic neuritis was first reported accompanying HAM . HTLV-1-Recurrent NMO associated with HTLV-1 infection has been reported , and basOverlap syndrome refers to HTLV-1-infected patients suffering more than one HTLV-1-associated inflammatory condition. HTLV-1 infection is considered to affect systemic immunological homeostasis, because T-cells are activated by HTLV-1 infection and cytokines and chemokines are produced from HTLV-1-infected cells. These alterations result in damage to resident cells in human. Clinically, overlap of HU and HAM is seen in less than 15% of cases . KCS andAccording to basic investigations, transgenic mice expressing HTLV-1 Tax develop inflammatory arthropathy, and transgenic rats expressing HTLV-1 env-pX develop Sjögren’s syndrome, arthropathy, vasculitis, and polymyositis . RegulatSince ATL-related ocular manifestations are rare and much less frequent than the prevalence of HU, manifestations have been documented in sporadic case reports or case series. Although several ocular manifestations have been identified, the frequencies remain unclear. Our recent nationwide survey in Japan revealed the frequency of ATL-related ocular manifestations against the background of the highest endemic rate among developed countries. According to the survey, the most frequent manifestation is intraocular infiltration, followed by opportunistic infection , KCS, and scleritis .Compared with other forms of leukemia, ATL cells are much more adept at infiltrating the eyes . PreviouInfiltration of ATL cells into the eye provides a specific sign, which we have termed the “knob-like ATL cell multiple ocular infiltration” (KAMOI) sign. At the ocular surface, this KAMOI sign can be seen most prominently at the bulbar conjunctiva around the corneal limbus and at the palpebral conjunctiva around each lacrimal punctum . KAMOI sAs for methods of imaging diagnosis, the slit-lamp scope and fundus scope have been useful to detect infiltrating lesions in the eye. OCT, one of the most recently developed modalities, is now able to identify ATL infiltration in detail . Recent As for laboratory diagnosis, methods for the analysis of ocular samples have evolved. Although cytology offers a precise method for detecting ATL cells, clinically obtainable ocular samples are limited in terms of quantity. PCR methods thus offer clinical advantages in ophthalmologic diagnosis . DetectiTherapy for intraocular infiltration previously relied on systemic ATL therapy such as intensive multi-agent chemotherapy , interfeAccording to our nationwide survey, the most frequent infectious manifestation among ATL patients was CMV retinitis ; Table 1As for imaging diagnosis, widefield fundus photography and OCT are helpful for identifying CMV retinitis, as these techniques can capture cytomegalic cell infiltration and accompanying retinal necrosis . DetectiOur nationwide survey identified co-infections with CMV such as herpes simplex virus and toxoplasma can be seen in some ATL patients . In addiSimultaneous development of ATL and ocular inflammation has been reported . InvasioIn AIU, ophthalmic examinations can detect infiltrating cells in the anterior chamber/vitreous, and retinal vasculitis. Diagnosis requires exclusion of ATL infiltration and opportunistic infections. Examination of ocular samples using PCR methods and cytology is useful. As for treatment, AIU often responds to topical steroid eye drops, but if the eye drops do not achieve adequate response, systemic steroids should be considered.ATL-related scleritis  has beenFor treatment, daclizumab  and denileukin diftitox (an immunotoxin fusion protein that targets IL-2R) reportedly improve scleritis in patients with underlying ATL . This suDrug-induced uveitis is a well-known phenomenon among ophthalmologists. Recently, a variety of drugs have been applied to HTLV-1 carriers, but the adverse effects of these drugs on the eye have not been fully characterized. Although HTLV-1 carriers face a credible threat of intraocular inflammation, safety assessments of drugs have not been completed and current guidelines do not mention whether patients should be checked for HTLV-1 infection prior to the administration of drugs.in vitro -blocking biologics were first introduced for inflammatory diseases, with other biologics such as IL-6-blocking agents introduced subsequently. Recently, we experienced a case in which a patient with RA and HTLV-1 infection received an IL-6-blocking agent, which reactivated HTLV-1 and induced HU in the eye . We havein vitro . FurtherAlthough great advances in ophthalmology have been seen since the discovery of associations between HTLV-1 and ocular diseases, various issues remain to be resolved. In terms of understanding the mechanisms involved, basic research into HTLV-1-associated diseases is still nowhere near complete, with remaining issues including how HTLV-1-infected cells break the BOB, and what roles HTLV-1-infected cells play in the eye. Although multiplex and broad-range PCR have been applied to HTLV-1-associated ocular diseases and the accuracy of diagnosis has vastly improved, diagnosis remains a complicated process requiring diagnosis by exclusion. In the future, advancing technologies such as flow cytometric analysis using ocular fluid  might prLooking back on the history of HTLV-1-associated ocular diseases, information accumulated from all over the world has helped to advance medical care for HTLV-1-associated ocular diseases since the discovery of the relationship between HTLV-1 and ocular diseases. However, HTLV-1-associated ocular diseases have been mainly reported from limited areas, and numbers of patients with HTLV-1-associated ocular diseases remain unclear for areas such as Africa and Australia. This may be due to low awareness among ophthalmologists in these areas regarding HTLV-1-associated ocular disease. In addition, HTLV-1-associated ocular diseases have now spread globally in conjunction with the migration of HTLV-1-infected individuals. An overview of these diseases in vast areas of the world is therefore needed, to provide information on HTLV-1-associated ocular diseases and prevent visual loss around the globe.As shown in this article, continued accumulation of knowledge from recent clinical and basic research has brought many new insights. Application of advances in ophthalmic technologies has brought about much more sophisticated medical care. Recent diagnostic technologies such as multiplex/broad-range PCR and OCT and treatment methods such as the direct approach through IVI of the drug have contributed to improvements in the diagnosis and treatment of HTLV-1-associated ocular diseases. In the future, development of techniques such as flow cytometry might represent the next step in advancing diagnostic methods, but the fact that only small-volume samples are able to be obtained from the eye must be addressed. In terms of treatment methods, development of new specific topical drugs based on the pathogenesis of HTLV-1-associated ocular diseases might be desirable, allowing direct injection into the eye. To provide forward-looking medical care for HTLV-1 in ophthalmology, various avenues of research must be explored to improve the quality and maintenance of vision among HTLV-1-infected patients. The constant effort of ophthalmologists is needed to achieve excellent vision for HTLV-1-infected patients.KK conceptualized and wrote the manuscript.The author declares that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Compartmentalization of diverse types of signaling molecules contributes to the precise coordination of signal propagation. The primary cilium fulfills this function by acting as a spatiotemporally confined sensory signaling platform. For the integrity of ciliary signaling, it is mandatory that the ciliary signaling pathways are constantly attuned by alterations in both oscillating small molecules and the presence or absence of their sensor/effector proteins. In this context, ciliary G protein-coupled receptor (GPCR) pathways participate in coordinating the mobilization of the diffusible second messenger molecule 3′,5′-cyclic adenosine monophosphate (cAMP). cAMP fluxes in the cilium are primarily sensed by protein kinase A (PKA) complexes, which are essential for the basal repression of Hedgehog (Hh) signaling. Here, we describe the dynamic properties of underlying signaling circuits, as well as strategies for second messenger compartmentalization. As an example, we summarize how receptor-guided cAMP-effector pathways control the off state of Hh signaling. We discuss the evidence that a macromolecular, ciliary-localized signaling complex, composed of the orphan GPCR Gpr161 and type I PKA holoenzymes, is involved in antagonizing Hh functions. Finally, we outline how ciliary cAMP-linked receptor pathways and cAMP-sensing signalosomes may become targets for more efficient combinatory therapy approaches to counteract dysregulation of Hh signaling. Compartmentalization is an important aspect of intracellular signaling, both in contributing to the precise coordination of signal propagation and in ensuring specificity of signaling outcomes ,2. PrimaOne central ciliary signaling component is the conserved and ubiquitous second messenger 3′, 5′-cyclic adenosine monophosphate (cAMP). First discovered by Sutherland and Rall in 1958, cAMP was proposed to act as a signal effector molecule downstream of hormones like epinephrine, dopamine, and prostaglandin . This reIntracellular cAMP mobilization is regulated by a family of adenylyl cyclases (ACs), which synthesize cAMP from adenosine triphosphate (ATP)  12]. Th. Th12]. Prime examples of cAMP-binding proteins are the functionally diverse type I and II holoenzyme complexes of PKA, isoforms of cAMP-dependent exchange proteins (Epac), cyclic nucleotide-gated ion channels (CNGs), and Popeye domain-containing proteins (POPDC) ,47,48,49The prototypical and macromolecular cAMP-effector protein complex is nucleated by the tetrameric PKA holoenzyme ,44. DiffAKAPs sequester PKA holoenzymes to specific subcellular nanodomains. The structures of PKA type I and type II holoenzyme complexes differ considerably in their functions and solved structures ,64,65,66Nearly every cell in the body assembles one primary non-motile cilium. The primary cilium is an antennae-like, microtubule-based protrusion from the apical membrane of most mammalian cell types . DynamicThe primary cilium is essential for this response . The corThe direct visualization of cAMP dynamics in the cilium has proved to be technically challenging, and measurements of resting cAMP levels in primary cilia vary ,92,93,94Interestingly, Gpr161 also contains a PKA consensus phosphorylation site, and has been shown to be a target for PKA phosphorylation . This suIt is an open question if the cAMP-sensing Gpr161/PKA-type I signalosome is indeed the central platform for integrating and orchestrating ciliary cAMP fluxes, compartmentalized type I PKA activities, Gli processing, and Hh counter-regulation. The next major progress for shedding light on the underlying mechanism of ciliary Gpr161 function will be accomplished upon the identification of Gpr161 ligands. In addition, detailed structural information of the organization of the Gpr161–signalosome complex will be essential for understanding ciliary compartmentalization and its role in the basal repression of Hh signaling .Recently, it has been shown that cAMP-induced PKA activities are not just central to Hh regulation. General cAMP fluxes have been linked to regulate cilium formation/removal. This is related to the recruitment of PKA and NIMA related kinase 10 (NEK10), a protein that is essential for ciliogenesis in vertebrates, to the pericentriolar matrix through the pericentriolar material protein 1 (PCM1). Thus, high cAMP levels lead to PKA-mediated phosphorylation of NEK10, which triggers its degradation and ultimately results in cilia resorption . InteresCilia host a collection of GPCRs that are coupled to Gαs or Gαi. The following reviews summarize the identity of ciliary localized GPCRs ,6,7 and Further, a macromolecular protein complex comprising the ciliary calcium channel polycystin-2, AC5/6, PKA, PDE4c, and the A-kinase anchoring protein AKAP150 has been identified within the primary cilia of renal epithelial cells . PolycysHh regulates developmental and physiological cell/tissue states by supervising cell growth, survival, fate, and patterning ,112. In The central role of PKA activity in the basal repression of Hh signaling is well documented. However, there is some ambiguity about how activation of Smo relieves the inhibitory effect on Gli-dependent transcriptional activation . The undLast, but not least, we envision that one specific GPCR may become an important candidate for antagonizing deregulated Hh signaling functions through cAMP mobilization. Gpr161 activities seem to be one of the long-sought factors for creating spatio-temporal controlled cAMP-gradients in the ciliary Hh signaling pathway . Smo act


Saccharomyces cerevisiae, proteins exist that are efficiently imported into peroxisomes despite having neither PTS1 nor PTS2 and for which no other import signal has been determined. An example of such a protein is S. cerevisiae acyl-CoA oxidase (AOx) encoded by the POX1 gene. While it is known that its import is driven by its interaction with the N-terminal segment of Pex5p, which is separate from its C-terminal PTS1-recognizing tetratricopeptide domain, to date, no AOx polypeptide region has been implicated as critical for this interaction, and thus would constitute the long-sought PTS3 signal. Using random mutagenesis combined with a two-hybrid screen, we identified single amino acid residues within the AOx polypeptide that are crucial for this interaction and for the peroxisomal import of this protein. Interestingly, while scattered throughout the primary sequence, these amino acids come close to each other within two domains of the folded AOx. Although the role of one or both of these regions as the PTS3 signal is not finally proven, our data indicate that the signal guiding AOx into peroxisomal matrix is not a linear sequence but a signal patch.The specificity of import of peroxisomal matrix proteins is dependent on the targeting signals encoded within their amino acid sequences. Two known import signals, peroxisomal targeting signal 1 (PTS1), positioned at the C-termini and PTS2 located close to N-termini of these proteins are recognized by the Pex5p and Pex7p receptors, respectively. However, in several yeast species, including  Saccharomyces cerevisiae. Few studies have been devoted to this protein, and our understanding of its import mechanism is limited. Nevertheless, these studies revealed that S. cerevisiae AOx is recognized by the Pex5p receptor, the same that functions in the PTS1-dependent import, but the recognition domain for AOx lies within the N-terminal part of Pex5p, separate from its C-terminal tetratricopeptide domain (TPR) that binds the PTS1 signal  or near the N-terminus (PTS2) of their polypeptides by receptors that recognize these signals, Pex5p or Pex7p, respectively . For thr1 signal . The reg1 signal . This su1 signal . Moreove1 signal . It remain vivo import tests demonstrated the importance of these amino acid residues for the import of AOx into the peroxisomal matrix, indicating that they form a signal patch that functions as novel peroxisomal targeting sequence.In this study, we used random mutagenesis of the AOx-encoding gene to identify AOx amino acid residues that are involved in its interaction with the N-terminal half of Pex5p and its import to peroxisomes. Surprisingly, the amino acid substitutions that affected AOx-Pex5p interaction and AOx import were scattered widely throughout the central part of the AOx polypeptide, comprising approximately half of the whole protein. However, in the model of the AOx tertiary structure, they cluster together into two small regions. The in vivo. The yeast strains were grown at 28°C either in rich YPD medium or in synthetic complete (SC) selective medium with 2% glucose or with a mixture of 0.1% glucose, 2% ethanol, 0.1% oleic acid, and 0.25% Tween 80 to induce peroxisome proliferation.The XL1-Blue (Stratagene) bacterial strain was used for plasmid construction and propagation. PJ69-4A  was usedThe plasmids that were used in this study are listed in POX1 ORF was randomly mutagenized by mutagenic PCR using a GeneMorph II random mutagenesis kit , according to the manufacturer’s protocol, to obtain an average single mutation per 1000 bp. Two hybrid libraries were made directly in yeast cells using gap repair method by transforming PCR products together with linearized pGBT9-AOx two-hybrid plasmid into a PJ69-4A strain previously transformed with the pGAD-Pex5p136–292 plasmid. Transformant clones displaying affected growth on the medium without adenine were isolated; plasmid DNA was prepared and amplified from them; and the inserts including the POX1 ORF were sequenced.POX1 ORF, selected in a two-hybrid screen, were transformed into the PJ69-4A strain together with the pGAD-Pex5p136–292 plasmid. The two-hybrid interaction displayed by the transformant clones was quantified by a plate test of growth in medium without adenine supplementation and by measuring the intracellular β-galactosidase activity according to the method published by pGBT-AOx plasmids bearing mutations within the 1 protein modeling software was used to model the structure of S. cerevisiae AOx based on its homology to AOx proteins in other organisms, for which crystallographic structures are known. The structures of acyl-coenzyme A oxidase in Yarrowia lipolytica , Rattus norvegicus , and Arabidopsis thaliana (1W07) were automatically selected as the best templates according to the combination of the Blast E-value, sequence coverage, and structure quality. For each template, up to five alternate alignments with the target sequence were used, and up to 50 different conformations were tested for each modeled loop. The resulting models were evaluated according to structural quality . The model with the highest score was built based on the 5YS9 structure; however, the chimerical model, in which some parts were adopted from other highly scored models , scored even higher  that direct ctor see , we founTo distinguish the recurring substitutions from the remaining ones, they were marked in S. cerevisiae AOx. While the structure of this protein has not yet been determined experimentally, the crystallographic structures of AOx proteins from many organisms belonging to other systematic groups are known. Importantly, two isoenzymes of Y. lipolytica AOx, ACOX1, and ACOX3, have been crystallized recently, and their structures were determined at 2.5 Å resolution  enabled the modeling of yeast AOx, based mostly on the template of YlACOX1 and YlACOX3 crystallographic structures, at a true atomic level of precision, particularly for the residues located in the core region of the protein. Yasara Structure package v. 19.5.5 was used for this purpose . The resulting model of the whole ScAOx molecule dimer and the enlargement of the region of interest is presented in The observation that the amino acid residues whose substitution affects the interaction of AOx with the Pex5p receptor are localized in several hot spots in the AOx polypeptide strongly suggested that these residues participate in this interaction. Therefore, we attempted to localize them within the natively folded AOx protein. To this end, we generated a structural model of solution , b. The in vivo import test, this substitution did not affect the import of AOx into peroxisomes. The representative images of cells expressing the wild-type AOx-GFP and those with substitutions found in clones 24, 40, 83, and 119 are shown in Amino acid residues participating in the AOx interaction with the N-terminal region of Pex5p are also likely to be important for AOx peroxisomal import. To test this assumption, the mutations in the clones that we selected in the two-hybrid screen were introduced into the gene encoding the full-length AOx C-terminally tagged with GFP, to determine their influence on the import of AOx into peroxisomes. The plasmids constructed this way and the plasmid bearing no mutations were separately transformed into BYpex5Δ,pox1Δ containing the mRFP-SKL plasmid encoding a peroxisomal fluorescent marker and pRS-Pex5pWT encoding the wild-type Pex5p receptor. For comparison, the strains bearing plasmids encoding the original AOX-GFP protein and pRS-Pex5pI264K encoding the mutated Pex5p receptor or not expressing Pex5p receptor at all were included in the analysis. Transformed cells were grown in oleate-containing medium to promote the proliferation of peroxisomes and examined by fluorescence microscopy. The cells were categorized according to the extent to which AOx-GFP was imported into the cells: perfect peroxisomal import, mixed peroxisomal/cytosolic localization and only cytosolic localization. The results of this experiment are shown in in vivo import of the wild-type AOx polypeptide fused to GFP in the presence of the mutated Pex5p receptor, in which isoleucine at position 264 was substituted with lysine (IK labeled column). As previously found, substituting this amino acid with lysine (For comparison, h lysine  or with h lysine  compromiS. cerevisiae peroxisomes has remained a mystery for decades. In contrast to our increasing understanding of the evolutionary conserved peroxisomal targeting of protein via PTS1- or PTS2-dependent pathways, our knowledge on the mechanisms of peroxisomal import of AOx and related proteins is still scarce. Based on the few known facts accumulated for peroxisomal targeting of these proteins we may conjecture that the corresponding mechanisms are distinct.The mechanism by which AOx is imported into Hansenula polymorpha alcohol oxidase (Y. lipolytica ACOX import is driven by its interaction with Pex20p, known as a coreceptor in the PTS2-dependent pathway, but it does not rely on this pathway or on the PTS1/Pex5p mechanism (Candida tropicalis AOx (S. cerevisiae catalase A (S. cerevisiae AOx, which is imported, as we have shown recently, independently of PTS1 and PTS2 (S. cerevisiae AOx with precision to individual amino acid residues. Using random mutagenesis of the POX1 ORF combined with the two-hybrid screen, we identified residues whose substitution abolishes the AOx interaction with Pex5p and makes AOx peroxisomal import less efficient. Remarkably, these amino acids, namely K159, N161, Q324, and perhaps also A333, cluster in a putative Pex5p-interacting domain that is, as one could expect, located at the surface of the deduced model of AOx. Thus the binding interface is formed by a cluster of several amino acid residues from two regions of the AOx polypeptide separated by 160 amino acids, which are in close proximity in the natively folded protein.The import of budding yeast AOx into peroxisomes is dependent on the known PTS1 receptor Pex5p, but does not involve its PTS1-recognizing TPR domains or the C-terminal amino acids of AOx. Instead, AOx binds to the N-terminal part of Pex5p , which l oxidase . On the echanism . Neverthalis AOx  and for talase A . The samand PTS2 . HoweverMoreover, we identified the FAD-binding pocket as another region, where amino acid substitutions affect AOx interactions with Pex5p and its peroxisomal import, albeit to a lesser extent. Notably the FAD-binding pocket, as well as residues substituted in clone 83, are buried inside the protein, as shown in the model of AOx structure see , 3.in vivo (see in vivo AOx-GFP import was studied in the presence of full-length Pex5p. This suggests that the import of AOx depends also on interaction of another AOx region, additional to K159, N161, Q324 domain, with the region of Pex5p outside its 136–292 domain. That is why we see some import of AOx-GFP proteins with substitutions preventing two-hybrid interaction (compare lower part of in vivo import of AOx are similar to the effect of I264K substitution in Pex5p on the interaction and in vivo import of wild-type AOx (compare the bottom of The two sets of data: two-hybrid interaction of AOx with Pex5p see  and AOx-vivo see  correlatin vivo import of AOx-GFP. It could also weaken or strengthen the interaction of AOx K159, N161, Q324 domain with 136–292 region of Pex5p receptor (compare upper part of The postulated additional Pex5p-interacting region in AOx may be the FAD-binding pocket as revealed by our two-hybrid screen. Alternatively, it may be located elsewhere and the substitutions within FAD pocket, as well as the substitutions N287K, G337D found in clone 83, may simply distort the folding of AOx. This would affect the in vivo import defect caused by Q324R substitution (compare columns labeled 82 and 119 in The inconsistency regarding clones 82 and 119 is more difficult to explain. Perhaps an additional substitution in clone 82 of D344 residue, buried inside the AOx molecule, distorts its structure in a certain way and thereby alleviates the S. cerevisiae AOx. Thus, our data revealed the matching partner to the prospective PTS3-recognizing domain within the N-terminal region of Pex5p, which was discovered almost two decades ago. Remarkably, the data indicate that the PTS3 is not a linear sequence as PTS1 and PTS2 but is composed of amino acids that are distant from each other in the primary sequence but are in close proximity in the folded protein. Thus, in contrast to the linear PTS1 and PTS2, the PTS3 is a signal patch.In summary, the surface-located domain composed of K159-N161 and Q324 amino acid residues is a very convincing candidate for the long-sought PTS3 import signal identified for H. polymorpha alcohol oxidase was also shown to abolish its peroxisomal import (The data presented in this paper provide the basis for further studies on the import mechanisms of AOx and similar proteins from other species. One open question concerns the structure of analogous PTS3 signals within Cat2p, Fox2p and Cta1p, the peroxisomal import of which can be driven exclusively or partially by their interaction with the 250–270 amino acid region of Pex5p. Another intriguing finding provoking further studies is the possible involvement of FAD-binding in AOx interaction with its receptor and in peroxisomal import. Notably, the amino acid substitution within the FAD-binding fold of the l import , indicatThe datasets generated for this study are available on request to the corresponding author.MS and RE: conceptualization and funding acquisition. BK, AC, JP, KK, RE, and MS: methodology. RE and MS: validation. MS: formal analysis, resources, data curation, visualization, supervision, and project administration. BK, AC, JP, KK, ŁR, ZF, WG, AS, and MS: investigation. ER and MS: writing – original draft, review, and editing.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The handling Editor declared a past collaboration with one of the authors RE.


Human T cell leukemia virus type 1 (HTLV-1) is a human retrovirus that is associated with two main diseases: HTLV-associated myelopathy/tropical spastic paraparesis (HAM/TSP) and adult T cell leukemia/lymphoma (ATL). Chemokines are highly specialized groups of cytokines that play important roles in organizing, trafficking, homing, and in the migration of immune cells to the bone marrow, lymphoid organs and sites of infection and inflammation. Aberrant expression or function of chemokines, or their receptors, has been linked to the protection against or susceptibility to specific infectious diseases, as well as increased the risk of autoimmune diseases and malignancy. Chemokines and their receptors participate in pathogenesis of HTLV-1 associated diseases from inflammation in the central nervous system (CNS) which occurs in cases of HAM/TSP to T cell immortalization and tissue infiltration observed in ATL patients. Chemokines represent viable effective prognostic biomarkers for HTLV-1-associated diseases which provide the early identification of high-risk, treatment possibilities and high-yielding clinical trials. This review focuses on the emerging roles of these molecules in the outcome of HTLV-1-associated diseases. Human T cell leukemia virus type 1 (HTLV-1) belongs to the Retroviridae family, the orthoretrovirinae subfamily and to the deltaretrovirus genus . HTLV-1 The exact mechanism of the development and pathogenesis of HAM/TSP and ATL among less than 3–5% of HTLV-1 carriers has not fully understood. However, it has been reported that genetic background such as human leukocyte antigen (HLA) and HTLV-1 genomic variation are associated with the risk of HAM/TSP among HTLV-1 carriers . The maj+ clones in HAM/TSP patients produce a wide variety of members of cyto/chemokine family, such as Interferon-gamma (IFN-γ), Tumor necrosis factor-alpha (TNF-α), Interleukin (IL)-2, Macrophage inflammatory protein 1-alpha (MIP-1α)/CCL3, beta (MIP-1β)/CCL4, and Matrix metalloproteinase-9 (MMP-9), which promote the inflammatory response  are important factors for the induction of chronic inflammation observed in the CNS of HAM/TSP patients . Plasma ATL is a peripheral T-cell malignancy with poor prognosis . In cont+CD25+ FOxp3low and express cytotoxic T lymphocyte-associated antigen (CTLA)-4 with no regulatory activity  which regulate migration and trafficking of various types of immune cells through interactions with a subset of cell surface seven-transmembrane G protein-coupled receptors (GPCRs) . To dateIn addition to their role in the immune system, chemokines also have a wide range of effects on many different cell types including various types of the CNS or endothelial cells, where they result in either angiogenesis or angiostasis . They arOn the basis of the arrangement of the two N-terminal cysteine residues in their biochemical structure, chemokines are further divided into four major subfamilies, CXC, CX3C, CC, and C, depending on whether the first two cysteine residues have either one or more amino acids between them  or are adjacent (CC) . CXC cheAccording to their function and pattern of expression, chemokines have also been classified into homeostatic and inflammatory groups or both. The homeostatic chemokines are constitutively expressed and important in many physiological processes and are involved in normal circulation and trafficking of leukocytes in lymph nodes and thymus, while the production of inflammatory group is induced by infection or other proinflammatory stimuli resulting in recruitment of leukocytes to the insulted regions, in particular, due to infection .Chemokine receptors are subdivided into CCR, CXCR, CX3CR, and CR based on the ligand to which they bind. The interesting fact about the chemokine receptors is the great level of redundancy observed in their effects on targets in which most of them can bind to several chemokines and most chemokines utilize more than one receptor . During HTLV-1 Tax and HBZ viral products are associated with oncogenic transformation and ATL pathogenesis . Tax is + and CD8+ T cells and B cells) but not in NK cells, monocytes, or granulocytes  from uninfected subjects , 2014. THBZ also plays a crucial role in the pathogenesis of ATL and HAM/TSP . It has HTLV-1 also incorporates viral protein Tax, and viral mRNAs including Tax, HBZ, and env, into exosomes derived from infected cells, which contribute to enhanced cellular FLICE-like inhibitory protein (cFLIP)-dependent survival of the recipient cell with induction of NF-kB, and resistance to apoptosis, by activation of AKT. Therefore, the exosomes derived from HTLV-1 infected cells might associate with HTLV-1 pathogenesis .+ CD25+ CCR4+ Foxp3– T cells which serve as reservoirs for HTLV-1  has been clearly investigated. It has been demonstrated that proteolipid protein-specific T-cell clones and production of the murine CCL1 homolog associates with EAE . CCL1 isCXCL11 or Interferon inducible T cell alpha chemoattractant (ITAC) enhances the chemotaxis of activated T cells and NK cells and its gene expression is induced by IFN-γ. CXCR3, the CXCL11 receptor, is also highly expressed on activated T cells, memory T cells, and NK cells and IFN-γ promotes its gene expression. It is also reported that CXCL11 is involved in the pathogenesis of cell mediated immunity . HAM/TSP+ T cells are selectively infected by HTLV-1 and converted into abnormal cells leading to the chronic inflammation in HAM/TSP patients /CCL2 mRNA in HTLV-1-infected T cell lines compared with uninfected cells have been reported . It is aThe interaction between stromal cell-derived factor 1 (SDF1)/CXCL12 and CXCR4 has been identified to play a paramount role in the migration of ATL cells . ExpressAnother chemokine receptor which is overexpressed in HTLV-1 infection is CXCR7 . CXCR7 sThe CXCL8 mRNA expression has been detected in peripheral leukemic cells of ATL patients and in cultured HTLV-1-infected T cell lines, including HUT-102, MT-1, SALT-3, and SKT-1B . CXCL8 pCCL18 is significantly expressed by DCs and mainly attracts naive T cells . Hence, The characteristic feature of ATL is organ infiltration by leukemic cells . ATL cel+) skin-homing memory T cells /CCL19 and secondary lymphoid-tissue chemokine (SLC)/CCL21 are well defined to play a role in the migration and homing of lymphocytes to lymphoid tissues via their common receptor, the CCR7 . It has lent ATL . Gain-oflent ATL .pCDNA3.1-TAX, is also thought to be able to induce CCR5 expression in the K562 cell line /CCL5 serves as a chemotactic factor for memory T cells that are produced by HTLV-1-infected cells and ATL cells in a Tax inducible manner . Althougell line . Furtherell line . The incCCL3/MIP-1α and CCL4/MIP-1β are highly produced in the culture supernatant and cytoplasmic fractions of tumor-infiltrating lymphocytes . It has Fractalkine or CX3CL1 is the only member of the CX3C chemokine subfamily . CX3CL1 It is well documented that ATL is associated with severe immunosuppression and patients with ATL are prone to opportunistic infections . HoweverCCL5 is highly chemotactic for memory T cells and monocytes . This chThe levels of some chemokines including CXCL9 and CXCL10 are undetectable in neutrophils from HTLV-1 infected and uninfected subjects . FurtherHTLV-1 infected individuals with overactive bladder, have similar serum concentrations of CXCL9 and CXCL10 compared to the carriers . Adding The following issues influence the limitation of the present review including: omission of relevant research, the authors’ point of view, and misconception in the translation of data from the primary literature. Controversial results of the study may have been observed due to an analysis of chemokine levels in serum/plasma, CSF and supernatant of HTLV-1 cell lines and PBMCs culture, sample size, disease duration, clinical status and heterogeneity of patients and HTLV-1 carriers. To the best of our knowledge, this review has been one of the first attempts to thoroughly explain the role of chemokines and their receptors in HTLV-1 infection; as such, a wide range of data regarding the chemokines and receptors are a strength of the study.In this study, the status of chemokines and chemokine receptors was discussed in HTLV-1 infection focusing on two major diseases associated with the virus: ATL and HAM/TSP. According to the available studies reviewed in this work, it is clear that chemokines play a critical role in inflammation and regulating the trafficking, migration, and tissue infiltration of leukocytes, and the dysregulation of various chemokines could play an important role in the pathophysiology of HTLV-1 associated disease. Clinical studies in HAM/TSP and ATL patients have been provided promising results to be hopeful to develop and recommend more effective inhibitors of chemokines and their receptors. Therefore, development of chemokine-directed therapeutic strategies for targeting leukocyte recruitment pathways could be a promising therapeutic approach for HTLV-1 infection.In this review, we have discussed the potential role of chemokines and their receptors in the outcome of HTLV-1 associated diseases. A great number of chemokines and their respective receptors have been implicated in the migration of HTLV-1 infected cells into CNS and skin lesions are a vital aspect in pathogenesis of HTLV-1-associated disease. Elevation of chemokines levels in serum and CSF in HTLV-1 infection has been considered as prognostic biomarkers and a promising surrogate marker for HAM/TSP treatment. The complexity of the chemokine network is perplexing. Thus, further investigation is required to target chemokine network that could open up a new door to therapy and prophylaxis against HAM/TSP and ATL.HR designed the study and wrote the manuscript. RZ designed and drafted the manuscript. MM drafted and wrote the manuscript. AM and GH revised the manuscript. ZV drafted the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


TSLP), inflammatory factors were detected by qRT-PCR. The injury level of liver was assessed via measurement of serum alanine aminotransferase (ALT) and serum aspartate aminotransferase (AST). H&E staining and Masson’s trichrome staining examine the liver fatty degeneration and fibrosis. MiR-142-5p and TSLP were differentially expressed and JAK-STAT signaling pathway was activated in the NASH group. Luciferase assay identified that TSLP was the downstream target of miR-142-5p. Through overexpression of miR-142-5p, ALT and AST in serum were inhibited, pro-inflammatory factors, liver fatty degeneration and fibrosis in liver tissues were decreased, while anti-inflammatory factors were increased. Overexpression of TSLP and JAK-STAT signaling pathway activation could reverse the effects of miR-142-5p on NASH. Taken together, overexpression of miR-142-5p could attenuate NASH progression via inhibiting TSLP and JAK-STAT pathway. MiR-142-5p might be a novel latent target for NASH therapy.This study aimed to figure out the underlying mechanism of miR-142-5p in the non-alcoholic steatohepatitis (NASH). Bioinformatics, luciferase assay and Western blot were performed. The NASH mouse model was established through feeding a high fat diet (HFD). Relative expressions of miR-142-5p, thymic stromal lymphopoietin ( Non-alcoholic steatohepatitis (NASH) is an increasing common chronic liver inflammatory disease characterized by hepatic steatosis and inflammation . It is dMicroRNAs (miRNAs), the cluster with 18-25 nucleotide RNAs, are one of the subgroups of non-coding RNAs. MiRNAs expression profiles differ between disease states and normal tissue, so it plays an important role in diseases . IncreasTSLP), which is a master regulator of Th2-driven inflammation, has been studied in various allergic diseases such as asthma and inflammatory bowel diseases (IBD). Park et al. reported that TSLP was a key mediator in the IBD progression [TSLP could trigger a cell-autonomous dendritic cell migration [TSLP might take an important part in the inflammatory diseases. Furthermore, a previous study has shown that TSLP induces the release of CCL2 from fibroblasts via STAT3, which in turn induces monocyte chemotaxis [Thymic stromal lymphopoietin    (P<0.05)  , 2B. Fro(P<0.05) . Furthered miRNA . RIP ass(P<0.01) , suggest(P<0.01) . We veri(P<0.05) . Dual-lu(P<0.05) .TSLP were evaluated. The Ldlr-/- mice fed with HFD (blank group) presented a decreased expression level of miR-142-5p compared with WT mice under CD (WT group). In NASH model, the mice treated with miR-142-5p agonist (A142 group) showed an upregulated expression of miR-142-5p compared with NC group (P<0.01) (The miR-142-5p expression was more markedly increased by mir-142 (16 mg/kg) than mir-142 (8 mg/kg) in the preliminary experiments (data not shown). Since, the mice in the A142 group were treated with 16 mg/kg mir-142. Wild-type C57BL/6 mice untreated (WT group) were used as control. Based on the established NASH mouse model, the expression of miR-142-5p and (P<0.01) . At the (P<0.01) . In acco(P<0.01) .P<0.05). However, this therapeutic effect was reversed by the agonist group and the overexpressed TSLP group (P < 0.05). Besides, miR-142-5p treatment group significantly inhibited the phosphorylation of the JAK-STAT signaling pathway. The use of activators and overexpressing TSLP groups reversed this inhibition. (P < 0.05).TNF-α, IFN-β, IL-6 and MCP-1, the pro-inflammatory factors were overexpressed in the Ldlr-/- mice fed with HFD compared with WT mice fed with CD. The anti-inflammatory factor IL-4 were decreased in the blank group compared with WT group. MiR-142-5p played an inhibiting inflammation role in NASH model. The expression of TNF-α, IFN-β, IL-6 and MCP-1 was all upregulated and IL-4 expression were downregulated in the group treated with miR-142-5p versus the NC group, and it could be rescued by colivelin or AAV/TSLP –4E. In aLP group . WesternTSLP, we detected the TGF-β and collagen-1α2 expression in miR-142-5p-colivelin group and miR-142-5p-TSLP group. The results revealed that TGF-β and collagen-1α2 expression was increased in A142-c group and A142-TSLP group compared with A142 group  in comparison with WT mice fed with CD (WT group). Besides, knockdown TSLP made no change to miR-142-5p expression but exhibited the inhibited expression of TSLP compared with NC group. Further, we investigated whether colivelin could influence the miR-142-5p or TSLP expression. The results showed that there was no relationship between colivelin and the expression of miR-142-5p and TSLP (MiR-142-5p expression was decreased and and TSLP .P<0.05), but the pathway activator group was able to reverse this therapeutic effect (P<0.05). This result indicated that JAK-STAT pathway was activated in the Ldlr-/- mice fed with HDF, suppressed in the NASH mice treated with AAV/sh-TSLP compared with NASH mice treated with sh-TSLP control and activated in the NASH mice treated with AAV/sh-TSLP and colivelin compared with sh-TSLP group.The expression of TNF-α, IFN-β, IL-6 and MCP-1 was over expressed and the expression of IL-4 was down regulated in NASH mouse model, indicating the high level of inflammation in NASH. Knockdown TSLP in NASH mice generally inhibited the pro-inflammatory factors expression levels and increased the anti-inflammatory factors expression, which suggested the inflammatory acceleration of TSLP in NASH. In addition, pro-inflammatory factors were upregulated and anti-inflammatory factors was down regulated in the NASH mice treated with AAV/sh-TSLP and colivelin compared with NASH mice only treated with AAV/sh-TSLP –7E. In ac effect . Furtherc effect , p-JAK1,As expected in the NASH model, the serum ALT and AST level, liver fatty degeneration, liver fibrosis and fibrosis-related genes expression were increased in the Ldlr-/- mice with HFD compared with WT mice with CD. As for the function of TSLP in NASH, we detected the above indicators and found that ALT and AST were secreted less, the fatty degeneration degree and fibrosis degree of liver tissue were attenuated and fibrosis-related gene expression were downregulated in the NASH mice treated with AAV/sh-TSLP compared with NC group. When we added colivelin into sh-TSLP group, the indicators as for NASH has been reversed compared with sh-TSLP group .NASH is a serious liver disease which can be defined by necro-inflammation, a variable amount of fibrosis and potential for progression to cirrhosis . Recent MiRNA reveals a biological function by affecting the expression of a target gene by incompletely binding to a target gene . MiRNAs TSLP encodes a hemopoietic cytokine and promotes T helper type 2 cells responses, which are linked to immune response in various inflammatory diseases, including asthma, allergic inflammation and chronic obstructive pulmonary disease. Li et al. found that TSLP promoted MRC-5 cells fibrosis and activated the expression of MAPK7, p-p38, p-ERK1 and p-JNK1 which are the biomarkers of MAPK pathway. The part of TSLP in MRC-5 cells indicated the potential therapeutic implication of TSLP in lung fibrosis of acute lung injury [TSLP was significantly overexpressed in liver tissue and serum of hepatitis C virus-related cryoglobulinemic vasculitis [TSLP in fibrotic liver tissue from chronic hepatitis C virus patients and detected the high level of TSLP expression [TSLP as regulator of inflammation and fibrosis in many diseases, the function of TSLP in NASH remains enigmatic. In this study, its high expression in NASH disease was also detected in the validation of animal models of disease. In addition, we further predicted the miRNAs regulated TSLP by miRanda and selected the miRNAs of this study by the selection of the miRNAs differentially expressed with the GEO database. MiR-326, miR-142-5p and miR-331-3p were the results after the intersection. Subsequently, only miR-142-5p was found to be statistically different in the detection of the three miRNAs in the NASH mouse model. Further dual luciferase reporter experiment revealed that the target gene of miR-142-5p was TSLP, so we believed that miR-142-5p might play a role in the occurrence of NASH by affecting the expression of TSLP. Similarly, a previous study showed that miR-142-5p can affect the expression of fibrotic genes, and fibrosis is one of the important causes of NASH [Currently, high-throughput technology has been identified as an effective means of detecting miRNAs that may be involved in the development of disease or have therapeutic potential in tissues or blood circulation disorders. In addition, luciferase reporter assays are gold markers for in vivo confirmation of pathologic miRNA/mRNA interactions , 29. Theg injury . Sansonnsculitis . Lee et pression . DespiteTSLP triggers inflammatory responses in the course of atopic diseases through JAK-STAT signaling pathway [TSLP on NASH and their effects on JAK-STAT signaling pathway were verified by establishing animal models of overexpression and down-regulation of miR-142-5p and TSLP, respectively.The KEGG analysis of NASH further showed that the JAK-STAT signaling pathway may play a role in NASH. This pathway have been elucidated to play a vital role in the signal transduction in the pathogenesis of many diseases . Accumul pathway . ConsistTSLP damage to NASH and whether it produced therapeutic effects. The results showed that overexpression of miR-142-5p resulted in a decrease in the expression of TSLP and a certain degree of reversal of liver damage caused by NASH, and the therapeutic effect of the experimental group after additional addition of TSLP was attenuated or even disappeared. Similarly, from the results, the therapeutic effect on NASH was significantly reduced by down-regulating the expression of TSLP. In the validation of the expression of JAK-STAT signaling pathway, we can see that when the expression level of miR-142-5p is increased, the expression of TSLP is decreased, and the expression of phosphorylated protein is decreased when the total protein expression is unchanged, thereby the activity of the JAK-STAT signaling pathway is inhibited. Consistent with our findings, it is well known that when the total protein is unchanged, the phosphorylation level of the corresponding protein is decreased or the phosphorylation level is decreased, the activity of inhibiting the signaling pathway is produced [TSLP is elevated, the JAK-STAT signaling pathway is activated, further aggravating the degree of liver damage in NASH. Therefore, we have reason to believe that miR-142-5p may affect the occurrence and development of NASH by regulating the expression of TSLP and then affecting JAK-STAT signaling pathway. This result is consistent with previous studies examining the relationship between JAK-STAT signaling pathway and NASH [ALT and AST are one of the important indicators reflecting the normal function of liver tissue, and its elevated content is often an important marker of liver function damage . TNF-α, produced . In addiand NASH .In summary, our study first proposed that miR-142-5p might be a protect factor in NASH. It could inhibit progression of NASH via TSLP/JAK/STAT axis, which provided a new potential therapeutic target of NASH. However, the limitation of this study lies in the fact that this part of the results is only proved by animal models and not in-depth discussion of its biological effect targets in combination with clinical investigation. Therefore, it is important to carry out appropriate clinical sample verification and in-depth mechanism exploration, and to provide evidence for the treatment of NASH through continuous in-depth research.http://www.ncbi.nlm.nih.gov/geo/). The dataset of GSE63067 was analyzed to visualize differentially expressed mRNAs, from which 7 human non-alcoholic steatohepatitis (NASH) and 7 healthy human liver tissues were chosen. The dataset of GSE33857 was used for aberrant expressed miRNAs. Differentially expressed genes (DEGs) as well as dysregulated pathways were uncovered through R packages and KEGG database. The target relationship between miRNA and mRNA was predicted by TargetScan 7.1 (http://www.targetscan.org/vert_71/). Differentially expressed mRNAs and miRNAs were screened out for further analysis with the criteria of |log2FC| > 1 and adj. P < 0.05 by Empirical Bayes method. The clustering analysis was based on “affy”, “affyPLM”, “RColorBrewer”, and “heatmap” packages.Microarray datasets GSE63067 and GSE33857 were obtained from Gene Expression Omnibus (GEO) database  were ligated downstream from the cytomegalovirus immediate early promoter within the gutted adeno-associated virus-2 (AAV) vector. We chose the CMV promoter for overexpression of TSLP plasmid  was constructed by Gene Chem Co. . While, a plasmid with U6 as a promoter was selected to interfere with the expression of TSLP  was constructed by Gene Chem Co. .10 PFU of each vector through tail intravenous injections. At last genomic copies of AAV (2 × 1010 to 2 × 1011) were used for tail-vein injections. Colivelin , a neuroprotective peptide and activator of STAT3, was used as an activator of JAK/STAT3 signaling pathway. At the age of 12 to 17 weeks old, mice were treated with 16 mg/kg mir-142, mir control, or PBS through tail intravenous injections (once a day for three days). The mir-142 and mir control were purchased from Genepharma . To activate JAK-STAT signaling pathway, mice were given 20 μg colivelin in 0.5ml physiological saline intraperitoneally for continuous 6 days.Animal-based experiments performed in this experiment were approved by Sichuan Academy of Medical Sciences and Sichuan Provincial People’s Hospital animal experiment ethics committee. The mice were all received human care according to the Guide for the Care and Use of Laboratory Animals . According to the predecessor’s NASH modeling method , all opeAll mice were sacrificed at 22 weeks by sedating them with 2.5% isofluorane. Mice blood was collected from inferior vena cava, centrifuged at 1500 g for 15 min, and stored it in −80°C. After that, livers were isolated and froze in liquid nitrogen quickly. Finally, we fixed the tissue in formalin and paraffin-embedded or stored it in −80°C.^-ΔΔCt method. Trizol reagent  were applied to extract the all the RNA of the cell or tissue fragment, then PrimeScript RT Reagent Kit  was employed for synthesis of cDNA under the instruction from manufacture. The miRNA was converted to cDNA using miScript cDNA synthesis kit  following the manufacturer’s protocol. Later, SYBR Select Master Mix and ABI Prism 7000 Sequence Detection System  were applied to carrying out qRT-PCR. β-actin and U6 were chosen as internal references for mRNA and miRNAs respectively. Relative expressions of mRNAs and miRNAs were quantified using 2A 3’-UTR wild type (wt) or mutant 3’-UTR (mut) of TSLP were constructed by PCR, and were then inserted into the multiple cloning sites in the pRL-TK Vector . Human 293T cells were cultured in six-well plate at the confluence of 70-80% and then were co-transfected with pRL-TK-TSLP-3’-UTR mut or pRL-TK-TSLP 3’-UTR wt plasmid in combination with 100 nM miR-142-5p or 100 mM negative control, respectively. Lipofectamine 2000 purchased from Invitrogen was used to transfect the vector. The luciferase activities after 48 h co-transfection were assessed by a Dual-Luciferase® Reporter assay system (Promega) and were normalized to firefly luciferase activity.RNA immunoprecipitation (RIP) assays were implemented using the Magna RIP RNA-binding Protein Immunoprecipitation Kit (Millipore), according to the manufacturer’s instructions. Anti-Ago2 antibody and normal IgG (Millipore) were used for immunoprecipitation. The co-precipitated RNAs were purified with phenol: chloroform: isoamyl alcohol and subsequently analyzed by qPCR to assess the enrichment of miR-142-5p and TSLP to Ago2.3 μm thick paraffin-embedded liver sections were stained with hematoxylin and eosin (H&E) and Masson’s trichrome to evaluate hepatic steatosis, inflammation, and liver fibrosis using an Olympus microscope . Hematoxylin and eosin staining were used for NAFLD activity score evaluation (NAS) . Masson’Olympus AU5400 automatic chemical analyzer (Olympus) was conducted to detect the levels of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST), which were reliable indicators of liver inflammatory injury, following the manufacturer’s instructions.Triglyceride assayTriglyceride (TG) levels were measured by a TG kit . The values obtained were normalized to total protein concentrations. Protein concentrations were measured using the BCA method.Frozen mouse liver tissue samples (about 50 mg) were homogenized in 650 mL of TNE buffer , 150 mmol/L NaCl) containing a phosphatase inhibitors and cocktail of protease . The incubation condition was 4°C for 45 min and the centrifugation condition was at 12 000 g for 5 min. BCA Protein Assay  was conducted to quantified protein content was quantified in the supernatant. Reducing sample buffer (Bio-Rad) was used to mix lysates for electrophoresis and subsequently the lysates were transferred onto polyvinylidene fluoride membranes (Thermo Fisher). Equal loading (50 mg) was verified using Ponceau red solution. Antibodies against TSLP (ab188766) was purchased from Abcam  and p-JAK1 (#3331S), JAK1 (#50994S), p-STAT1 (#9167S), STAT1 (#14994S), p-STAT3 (#52075S), STAT3 (#9139S) and anti-β-actin (#3700S) were obtained from Cell Signaling Technology . After incubating with HRP-labeled goat anti-rabbit IgG (H+L) , the Clarity Western ECl Substrate  was conducted to proceed immunodetection and the Las-4000 Imaging System and Image MultiGauge software  were used to reveal bands.All the statistical analyses were performed by GraphPad Prism (vesion6.0) software. The form of means ± standard deviation (SD) was used for the whole results. Student t-test (unpaired t-test) was used to compare means between differentiated and undifferentiated groups. Differences among the three groups were analyzed via one-way analysis of variance (ANOVA), Tukey’s test was performed in all pairwise comparisons. P values < 0.05 were considered as statistically significant.All procedures performed in studies involving animals were in accordance with the ethical standards of the Sichuan Academy of Medical Sciences and Sichuan Provincial People’s Hospital.Supplementary Figure 1Supplementary Table 1


Mepolizumab (MPZ), an anti-interleukin-5 antibody, is effective for the treatment of eosinophilic granulomatosis with polyangiitis (EGPA). However, its effectiveness has not been adequately evaluated in real-world clinical practice. In this study, we assessed the effectiveness and safety of MPZ (300 mg) for relapsing/refractory EGPA resistant to corticosteroids (CS) for 1 year in real-world settings.We administered MPZ (300 mg) to 16 patients with relapsing/refractory EGPA resistant to CS (Post-MPZ). We also retrospectively collected data from the same patients for the 12 months before the administration of MPZ (Pre-MPZ). The primary endpoint was the 12-month remission rate after MPZ administration and the secondary endpoints were the Birmingham vasculitis activity score (BVAS), vasculitis damage index (VDI), eosinophil counts, changes in concomitant CS doses/concomitant immunosuppressant use, MPZ retention rate, and incidence of adverse events. The clinical course was compared between Pre-MPZ and Post-MPZ.The 12-month remission rate after the initiation of MPZ was 75%. No change was observed in BVAS, eosinophil count, or concomitant CS dose over time in the Pre-MPZ group, whereas all these parameters were significantly decreased over time in the Post-MPZ group. The number of patients using concomitant immunosuppressant also decreased over time in the Post-MPZ group. VDI did not increase in either group. The MPZ retention rate was 100% and only three patients (18.8%) had infections. Changes in BVAS, eosinophil count, and cumulative concomitant CS dose were significantly lower in the Post-MPZ group than in the Pre-MPZ group. There was no significant difference in the changes in VDI between the groups.This study demonstrated that MPZ is effective and safe for EGPA. Furthermore, MPZ decreases disease activity, increases remission rate, and has a CS-sparing effect. MPZ is safe for the treatment of EGPA in real-world clinical practice.Comparing to Pre-MPZ, MPZ possesses the high remission rate and CS sparing effect.Eosinophilic granulomatosis with polyangiitis (EGPA) is a disease that is preceded by asthma or allergic rhinitis. EGPA causes various symptoms owing to vasculitis, including fever and purpura, and increases peripheral eosinophil counts , 2. CortIn recent years, mepolizumab (MPZ), an anti-interleukin-5 (IL-5) monoclonal antibody, has been reported to extend the remission period of EGPA and reduce the CS dose required . AlthougIn this study, MPZ (300 mg) was administered to 16 patients with relapsing or refractory EGPA who were receiving the standard of care, mainly CS , 8. In JThe patients were followed up for 12 months after the introduction of MPZ at our hospital and affiliated institutions, during the period between the domestic introduction of MPZ in May 2018 until August 2020 (Post-MPZ). Additionally, we retrospectively collected data from the same patients for the 12 months before the initiation of MPZ therapy (Pre-MPZ). All patients received maintenance therapy according to the standard of care. The standard of care in this study was defined as treatment with CS, intravenous CY, intravenous immunoglobulin, azathioprine, MTX, or CsA/TAC. The Human Ethics Review Committee of our university reviewed and approved this study (No. H27-014). We also complied with the Declaration of Helsinki. All participants provided informed consent prior to inclusion in the study. Details that might disclose the identity of the study subjects were omitted.This study was a multicenter and ambispective cohort study in which MPZ at a dose of 300 mg/month was administered to 16 patients with relapsing or refractory EGPA to investigate the effectiveness and safety of MPZ over 1 year.The primary endpoint was the remission rate. The secondary endpoints were the BVAS , vasculitis damage index (VDI)  , 12, eosThe serum concentration of IL-5 at the baseline (before MPZ initiation) was measured using the enzyme-linked immunosorbent assay (ELISA)  and compared with that of six age- and sex-matched healthy controls.U test.Data are expressed as the median (interquartile range). For statistical analysis, data from cases in which MPZ was discontinued or the disease relapsed were complemented using the last observation carried forward method. Differences between groups (Post-MPZ vs. Pre-MPZ) and between data measured at the baseline and each observation point  were compared using the Wilcoxon signed-rank test or McNemar test. Differences in the serum IL-5 concentration between the patients and healthy controls were compared using the Mann-Whitney P values are two-sided. Remission was defined as a BVAS score of 0 and CS less than 4 mg/day. All analyses were conducted using JMP version 14.0.0 (SAS Institute Inc.). The post hoc power of this study for the comparison between month 0 and the other observation points in the Post-MPZ group was 0.37 for BVAS, 0.99 for VDI, 0.36 for absolute eosinophil count, and 0.76 for concomitant CS dose .The timeframes of both groups were represented synchronously and compared [− 12 months (baseline) in the Pre-MPZ corresponded to 0 months (baseline) in the Post-MPZ group]. All reported P = *0.0063 using Mann-Whitney U test; Supplementary Fig. The characteristics of the patients are shown in Table The remission rates (the primary endpoint) were 6.3% (1/16 patients) at month 1, 12.5% (2/16 patients) at month 3, 6.3% (1/16 patients) at month 6, and 0% at month 12 in the Pre-MPZ group. The corresponding rates in the Post-MPZ group were 12.5% (2/16 patients) at month 1, 31.3% (5/16 patients) at month 3, 50.0% (8/16 patients) at month 6, and 75.0% (12/16 patients) at month 12. In this group, the remission rate increased at month 12 . P values were determined by Mann-Whitney’s U test. p* < 0.01: EGPA group (n = 16) vs. HC (n = 6).


Major signaling pathways, such as Notch, Hedgehog (Hh), Wnt/β-catenin and Hippo, are targeted by a plethora of physiological and pathological stimuli, ultimately resulting in the modulation of genes that act coordinately to establish specific biological processes. Many biological programs are strictly controlled by the assembly of multiprotein complexes into the nucleus, where a regulated recruitment of specific transcription factors and coactivators on gene promoter region leads to different transcriptional outcomes. MAML1 results to be a versatile coactivator, able to set up synergistic interlinking with pivotal signaling cascades and able to coordinate the network of cross-talking pathways. Accordingly, despite its original identification as a component of the Notch signaling pathway, several recent reports suggest a more articulated role for MAML1 protein, showing that it is able to sustain/empower Wnt/β-catenin, Hh and Hippo pathways, in a Notch-independent manner. For this reason, MAML1 may be associated to a molecular “switch”, with the function to control the activation of major signaling pathways, triggering in this way critical biological processes during embryonic and post-natal life. In this review, we summarize the current knowledge about the pleiotropic role played by MAML proteins, in particular MAML1, and we recapitulate how it takes part actively in physiological and pathological signaling networks. On this point, we also discuss the contribution of MAML proteins to malignant transformation. Accordingly, genetic alterations or impaired expression of MAML proteins may lead to a deregulated crosstalk among the pathways, culminating in a series of pathological disorders, including cancer development. Given their central role, a better knowledge of the molecular mechanisms that regulate the interplay of MAML proteins with several signaling pathways involved in tumorigenesis may open up novel opportunities for an attractive molecular targeted anticancer therapy. Developmental signaling pathways, such as Notch, Wnt/β-catenin, Hedgehog (Hh), and Hippo, are highly conserved in multicellular organisms. These signaling cascades play a significant role in embryonic development, to establish body plan and they are also involved in the onset/progression of several cancers. In eukaryotic cells, extracellular and intracellular signals can stimulate these pathways, which convert the stimuli in specific transcriptional events inside the cell. The transcriptional activation of eukaryotic genes is a multistep process, controlled by tightly regulated assembly of multiprotein complexes, constituted by transcription factors and co-regulators proteins (coactivators or co repressors) recruited on specific enhancers and promoters to determine the final outcome . The traMAML genes may be regulated by microRNAs and how some DNA viruses can modulate the expression of MAML proteins, thus sustaining viral carcinogenesis. Altogether, these events suggest that several controls are required to regulate the expression/activity of MAML proteins, being involved in several physiological or pathological processes.In this review, we focus our attention on Mastermind-like (MAML) transcriptional coactivators family, firstly described as integral part of the Notch signaling pathway , and nowThe Notch pathway is an evolutionarily conserved and finely orchestrated signal transduction, responsible for cell fate determination, embryonic patterning and development, in response to specific cues . The intDrosophila, LIN-12, and GPL-1 in Caenorhabditis elegans and Notch1-4 in mammals) occurs upon binding to specific ligands , expressed on neighboring cells. This occurrence determines sequential proteolytic cleavage events, sustained by ADAM  and presenilin (PS)/γ-secretase complex, which allow the release of the intracellular domain of Notch receptor (ICN), able to move into the nucleus and to interact with CSL  in Drosophila, and LAG-1 in C. elegans) a DNA-binding transcription factor (Hairy/Enhancer of Split genes , which encode a family of basic helix-loop-helix (bHLH) transcriptional repressors  is a neurogenic gene genetically associated with Notch function and isolated in early 1980 . The coactivator p300 interacts with MAML1 through a proline-rich motif (PXPAAPAP)  , at resiXPAAPAP) . Recent get gene . Altogetget gene .In addition, the crucial role of Mastermind1 in driving the Notch transcriptional activation is demonstrated by using MAML1 dominant negative mutants (DNMAML1), without the N-terminal domain. These variant fragments are able to inactivate the ICN-CSL-MAML ternary complex, by competing with the wild-type forms, thus allowing Notch-signaling to be switched-off .Likewise, glycogen synthase kinase 3 β (GSK3β) directly interacts with MAML1 N-terminus, but it is able to decrease Notch transcriptional activity, by inhibiting MAML1-dependent histone acetylation .Interesting is the observation that the TAD1 domain mediates also the interaction of MAML1 with components of other pathways, such as MEF2C , p53 Zh, and EGRIn addition, the MAML1 TAD2 domain, placed at the C-terminal region , contains glutamine-rich sequences that govern the MAML1 transcriptional activity . It is nThe mammalian Mastermind proteins are widely expressed in embryonic and adult tissues following a differential spatio/temporal distribution and showing a different affinity/strength to interact with the Notch receptors . Accordi217 and Lys299) of Mastermind1, is able to enhance its interaction with HDAC7, which in turn represses the transcriptional activity of MAML1 itself , is generated . SAHM1 i in vivo . Moreove in vivo .Firstly identified in the canonical Notch signaling pathway, MAML1 is now recognized as integral component of the main signal transduction cascades, as Wnt/β-Catenin, Sonic Hh and Hippo pathways. In all of these signaling pathways, MAML1 shows a strong ability to convert a plethora of stimuli in several biological processes, such as proliferation, differentiation, tissue development, and tumorigenesis, by modulating the expression of specific target genes.The Wnt/β-catenin signaling is an evolutionarily conserved pathway, vital for the development of living organisms, being able to modulate important processes, such as differentiation and proliferation . The actIn the off-state, β-catenin is localized in the cytosol and prone to proteasomal degradation through β-TrCP activity . Wnt procyclin D1 and c-Myc, important β-catenin effectors , Indian hedgehog (IHh), and Desert hedgehog (DHh). The three genes encode for three different proteins with specific functions: SHh plays a key role in the nervous system; IHh takes part in endoderm and bone tissue development and DHh controls the spermatogenesis  is an evolutionary conserved pathway that controls several physiological processes, such as embryonic development, tissue differentiation, cell growth and maintenance of stem cells . Firstly the fly . In mammogenesis .gli1 and ptch1) and genes involved in important cellular processes, as proliferation, survival and differentiation   to the Patched (Ptch1) receptor, characterized by twelve transmembrane spanning-domain . The lignd bcl2) . In partnd bcl2) . Persistnd bcl2)  of Hh sind bcl2) , and tumnd bcl2) . Recent la ovary  and in cla ovary .Drosophila ovary  in la ovary . More rela ovary . Cerebella ovary . The cerla ovary . It is kla ovary . Impairela ovary . Bio-infla ovary . Accordise model  with a nse model  and suggin silico analysis. Given the above observations, MAML1 behaves as a potent transcriptional coactivator of GLI1 to strongly empower the expression of specific SHh target genes, suggesting that MAML1 may be considered as a novel therapeutic target for developing innovative cancer treatment to contrast cancer growth and proliferation  gene family, key transcription factors involved in myogenic program during muscle differentiation. MEF2 proteins belong to the MADS box family, characterized by a DNA binding domain, the MADS-box, in the N-terminal region . Upon activation of the Notch signaling, caused by muscle injury, MAML1 switches its role to transcriptional coactivator for Notch, resulting in the expression of the Notch specific target genes, functioning as a molecule with a functional switch property  is expressed in response to various extracellular signals and diverse stress stimuli. Stimulation by growth factors rapidly induces expression of EGR1, which subsequently leads to the activation of downstream growth pathways . EGR1 acarcinoma .via a RelA/MAML1 functional complex. Indeed, when co-expressed with MAML1, RelA switches its subcellular localization, from the cytoplasm to the nuclear bodies. The TAD2 domain, located at the C-terminus of MAML1 is involved in the binding with NF-κB  family consists of five transcription factors, p50, p52, p65 (RelA), c-Rel, and RelB, which couple with each other to form hetero- and/or homo-dimers . NF-κB rth NF-κB . Notablytoxicity . These dtoxicity .in vitro. The analysis of –/–MAML1 mice long bones reveals a smaller mineralized region, when compared to the wild type mice. Moreover, histological sections of the area of primary spongiosa of the femoral diaphysis suggest skeletal defects in MAML1-null mice, due to an impairment of chondrocyte maturation. Interestingly, the inhibition of the Notch pathway do not affect MAML1-dependent Runx2 transcription, suggesting a Notch-independent role for MAML1 in Runx2 activity in bone development (Runt-related transcription factor 2 (Runx2) is involved in osteoblast differentiation and chondrocyte maturation during bone development. Runx2 belongs to the Runx transcription factor family and is characterized by a DNA-binding runt domain involved in the transcription of target genes . Differeelopment .t  chromosomal translocation, resulting in a fusion transcript containing the exon 1 of the N-terminus of the MEC translocated 1 gene (MECT1) with the transactivation domain of MAML2 (corresponding to exons two through five)  (q21q23), in secondary acute myeloid leukemia (AML) and in myelodysplastic syndrome (MDS). In the chimeric protein, the exon seven of MLL is fused to exon two of MAML2. The MLL-MAML2 fusion protein contributes to carcinogenesis in AML and MDS, by disrupting the Notch signaling pathway  of the salivary and bronchial glands is a recurrent gh five) . The MECgh five) , leadinggh five) . MECT1-Ms tumors  and in cs tumors . Likewis pathway . Interesn cancer . Interese tumors . YAP1-MAe tumors . Interese tumors .t  leads to fusion transcript that associates exon 1–7 of PAX3 with exon 2–5 of MAML3. The chimeric protein works as a potent transactivator of PAX3 response elements with a forced expression of target genes involved in neuroectodermal and myogenic differentiation , a drug largely used in neuroblastoma treatment, promoting hyperproliferation of tumoral cells . A novelntiation .In addition, an aberrant expression of MAML1 is described in esophageal squamous cell carcinoma (ESCC), human HCC and in head and neck squamous cell carcinoma, where it correlates with the clinicopathological features of tumors, predicting poor prognosis . Recentlin silico analysis reveals the presence of some MAML1 mutations in several cancer cell lines. These aberrations are mainly represented by nonsense and frameshift mutations and they could affect both the stability and the folding of MAML1 with functional consequences for Notch signaling  and by β-human papillomavirus (β-HPV), through the binding mediated by the acid LXXLL α-helical motif, located at the C-terminal acid domain . E6 protNotch, Wnt/β-catenin, Hh and Hippo are essential transduction pathways that play pleiotropic roles during vertebrate development and their deregulation may sustain the process of tumorigenesis. Each of these major pathways responds to distinct external/internal cellular stimuli that result in a coordinated set of molecular events, whose endpoint is a tight regulation of gene transcription. The broadly accepted paradigm of transcription regulation in eukaryotic cell is a multistep process, strictly controlled by a concerted action of transcription factors and coactivators that build protein assemblies, following a precise sequence for mediating gene activation.Despite the original identification of MAML1 as a component of Notch signaling pathway, MAML1 may be considered as “master regulator,” showing the ability to integrate different signaling pathways and to activate distinct biological programs in different tissues. Accordingly, MAML1 may activate several other co-regulators inside the canonical Notch signaling, such as p300 and CycC:CDK8, to amplify the signaling process, or interact with components of other signaling pathways, as GLI1/2, β-catenin, and YAP/TAZ . Indeed,SZ and MP conduced the systematic literature search, wrote the manuscript, and prepared the figures. FN contributed to the literature search and prepared the manuscript. IS and MF reviewed draft of the manuscript. DB put forward the idea of the manuscript and helped with editing. All authors contributed to the article and approved the submitted version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.