TECHNICAL FIELD

The present disclosure is directed to neonatal Neutrophil Inhibitory Factor (nNIF) and nNIF-Related Peptides (NRPs). The present disclosure is also directed to methods of using nNIF and NRPs for the inhibition of neutrophil extracellular trap (NET) formation. Furthermore, nNIF and NRPs can be used for the treatment of and the prophylaxis against inflammatory disorders and cancer BACKGROUND

Formation of neutrophil extracellular traps (NETs) can be an important component in the defensive armamentarium of neutrophils (polymorphonuclear leukocytes; PMNs) that allows them to capture, immobilize, and putatively kill microbes in the extracellular space (see Sorensen O E, et al.,Journal of clinical investigation.2016; 126 (5):1612-20, Brinkmann V, et al.,J Cell Biol.2012; 198 (5):773-83; Yipp B G et al.,Blood.2013; 122 (16):27B4-94; and Brinkmann V, et al.,Science.2004:303 (6663). 1632-5). NET formation occurs by a novel cell death process often called NETosis, although “vital” NETosis, in which the neutrophils do not immediately die, has also been described (see Yipp B G, et al.,Blood.2013; 122 (16):2784-94 and Yipp B G, et al.,Nature medicine2012; 18 (9) 1386-93). The molecular mechanisms leading to NET formation have not been completely dissected and may depend in pan on the stimulus (see Sorensen O E, et al.,Journal of clinical investigation2016; 126 (5):1612-20; Brinkmann V, et alJ Cell Biol.2012; 198 (5):773-83; Yipp B G. et al.,Blood.2013; 122 (16):2784-94; and Papayannopoulos V, et al.,J Cell Biol2010; 191 (3):677-91) Nevertheless, decondensation of chromatin and extrusion of DNA together with histones and granule contents are central events (see Sorensen O E, et al.,Journal of clinical investigation.2016; 126 (5):1612-20, Brinkmann V, et al.,J Cell Biol.2012; 198 (5):773-B3; Yipp B G, et al,Blood2013; 122 (16):2784-94; Yipp B G. et al.,Nature medicine.2012; 18 (9) 1386-93; Papayannopoulos V, et al.,J Cell Biol.2010; 191 (3):677-91). Deimination of histones mediated by peptidyl arginine deiminase 4 (PAD4) (see Wang Y, et al.,J Cell Biol.2009; 184 (2):205-13 Li P, et alJ Exp Med.2010; 207 (9): 1853-62; and Kolaczkowska E. et al.Nature communications2015; 6 (6673)) is thought to be a sine qua non for nuclear decondensation and NET formation (see Sorensen O E, et al.,Journal of clinical investigation.2016; 126 (5): 1612-20).

NET-mediated capture and elimination of pathogens may complement traditional PMN antimicrobial activities including phagocytosis and intracellular killing (see Brinkmann V, et al.,J Cell Biol.2012; 198 (5):773-83 and Nauseef W M,Immunol Rev.2007; 219 (88-102)). Clinical observations indicate that defects in NET formation contribute to intractable infections in some instances (see Brinkmann V; et al.,J Cell Biol.2012; 198 (5):773-83 and Bianchi M et al.,Blood2009:114 (13):2619-22), but the importance of NETs in pathogen killing in vivo remains unclear (see Sorensen O E. et al.,Journal of clinical investigation.2016; 126 (5): 1612-20, Brinkmann V. et al.,J Cell Biol.2012; 198 (5) 773-83, and Yipp B G, et al.,Blood.2013; 122 (16) 2784-94). Conversely, there is substantial evidence that NETs and NET-associated factors, including histones and granule proteases, mediate vascular and tissue injury and that NET-mediated injury is a previously-unrecognized mechanism of innate immune collateral damage to the host (see Sorensen O E, et al.Journal of clinical investigation.2016; 126 (5) 1612-20; Brinkmann V, et al,J Cell Biol.2012; 198 (5) 773-83, Yipp B G. et al.,Blood.2013; 122 (16):2784-94; Kolaczkowska E. et al.Nature communications2015; 6 (6673), and Xu J, et al.Nature medicine2009; 15 (11):1318-21). Experimental models and limited clinical observations suggest that intra- or extravascular NET formation contributes to tissue injury in bacteremia (Kolaczkowska E, et al,Nature communications.2015; 6 (6673); Clark S R, et al.,Nature medicine2007; 13 (4):463-9; and McDonald B, et al.Cell host&microbe2012; 12 (3):32433), transfusion-related acute lung injury (see Caudrillier A. et al.,J Clin Invest.2012; 122 (7) 2661-71), primary graft dysfunction after lung transplantation (see Sayah D M, et al.,American journal of respiratory and critical care medicine.2015; 191 (4) 455-63), sterile vasculopathies and immune inflammation (see Chen G. et al,Blood2014; 123 (24) 3818-27; and Lood C, et al.,Nature medicine.2016; 22 (2): 146-53), thrombosis (see Fuchs T A, et al.,Proc Natl Acad Sci USA.2010; 107 (36): 15880-5), and influenza (see Pillai P S, et al.Science(New York, N.Y.). 2016; 352 (6284):463-6). Thus, NET formation may be an important maladaptive activity of neutrophils (see Sorensen O E, et al.,Journal of clinical investigation2016; 126 (5):1612-20) if it is triggered inappropriately or is unregulated in infection and inflammation.

Human neonates have unique and complicated immune regulation, susceptibility to infection, and inflammatory pathology. Although the infant is in a sterile environment in utero, it can be challenged by pathogens and their products before or during labor (see McDonagh S, et al.,Journal of infectious diseases.2004; 190 (4) 826-34). Furthermore, newborns are rapidly colonized with bacteria after delivery a process associated with increases in circulating and bone marrow neutrophils (see Palmer C, et al.PLoS biology.2007; 5 (7):e177. Jost T, et al.,PloS one.2012; 7 (8):e44595, and Deshmukh H S, et al.,Nat Med.2014; 20 (5):524-30). Complex adaptations appear to have evolved that prevent excessive, injurious inflammation in the perinatal period and in the abrupt neonatal transition from the protected intrauterine environment to continuous microbial colonization and exposure (see Dowling D J, et al.Trends in immunology.2014; 35 (7):299310; Adkins B.,Immunologic research.2013; 57 (1-3):246-57, and Elahi S, et al.,Nature.2013; 504 (7478). 158-62). These adaptations may, however, be accompanied by increased susceptibility to infection (see Adkins B.,Immunologic research.2013; 57 (1-3) 246-57 and Elahi S, et al,Nature.2013; 504 (747B) 158-62). It has been found that PMNs isolated from umbilical cord blood of preterm and term infants do not form NETs when stimulated and have a defect in NET-mediated bacterial killing, suggesting such an adaptation (see Yost C C, et al.,Blood2009; 113 (25) 6419-27). Other investigators subsequently reported temporally delayed NET formation when isolated neonatal neutrophils were stimulated in vitro (see Marcos V, et alBlood.2009; 114 (23):4908-11, author reply 11-2).

