Our goal is to analyze the molecular mechanisms that underlie the formation of tubulointerstitial (TI) fibrosis, to devise more effective therapies to prevent its progression. We study the collagen receptor integrins 11 and showed that it downregulates collagen synthesis, and its loss leads to increased glomerular fibrosis following injury. Integrin 11 plays an anti-fibrotic action by negatively regulating the phosphorylation state of pro-fibrotic growth factor receptors via activation of the tyrosine phosphatase TCPTP. We have also found that integrin 11 is a negative regulator of TI fibrosis, as integrin 1KO mice show increased unilateral ureteral obstruction-induced fibrosis. Moreover, integrin 1KO collecting duct (CD) cells have increased activation of TGF- receptor (TR)-dependent pro-fibrotic signaling, such as phosphorylated Smad3 and collagen levels. TGF- exerts its functions via activation of the serine/threonine kinases TRI and TRII. Binding of TGF- to TRII leads to phosphorylation of TRI and subsequent activation of the major pro-fibrotic mediator Smad3. Interestingly, the cytoplasmic tail of TRII can also be phosphorylated on tyrosine residues. However, whether these tyrosines play any physiological or pathological role in renal cells is unknown. The novelty of this proposal is that integrin 1KO CD cells show increased basal levels of tyrosine phosphorylated TRII. This result suggests that, in renal cells, integrin 11 crosstalks with TRII and it might prevent its pro-fibrotic action by downregulating its tyrosine phosphorylation levels. Interestingly, inhibition of TCPTP in CD cells leads to increased Smad3 activation and collagen synthesis. This result, together with the finding that 3 tyrosines in the TRII tail can be potential substrates of TCPTP, forms the hypothesis that integrin 11 negatively regulates TRII tyrosine phosphorylation via activation of TCPTP. Thus, we propose that integrin 11/TCPTP-mediated dephosphorylation of TRII represents an important, but previously undescribed mechanism to selectively reduce TRII activation and consequent progression of fibrosis. To test this hypothesis:
Aim 1 will analyze the role of TRII-mediated pro-fibrotic signaling in TI injury in the integrin 1KO mice. We will cross integrin 1KO mice with global null or floxed Smad3 mice to determine if preventing TR/Smad3 axis in the collecting system is sufficient to ameliorate TI fibrosis in the 1KO mice. We will then determine if in vivo activation of TCPTP is beneficial in the setting of TI injury by counteracting TR-mediated pro-fibrotic action.
Aim 2 will determine the mechanisms whereby integrin 11 negatively regulates TRII. We will use in vitro approaches to analyze i) if TCPTP directly binds and dephosphorylates TRII; ii) if tyrosine residues are important to control TRII-mediated Smad3 activation and collagen synthesis; and iii) which tyrosine(s) controls TRII-mediated functions. This study will lead to the identification of a novel crosstalk between integrin 11 an TRII and, most importantly, a novel mechanism whereby TRII-mediated pro-fibrotic signaling can be negatively modulated.
Transforming growth factor-? (TGF-?) is widely recognized as one of the key mediators of renal fibrosis leading to chronic kidney disease (CKD); however anti-TGF-? therapy is not effective for the treatment of CKD because 1) TGF-? also exerts beneficial effects; and 2) activation of TGF-? downstream pro-fibrotic signaling can be TGF-? ligand independent. In this proposal, we provide evidence that in the collecting system of the kidney, the collagen receptor integrin ?1?1 crosstalks with the TGF-? receptor II (T?RII) and plays an anti-fibrotic role by negatively regulating T?RII-mediated pro-fibrotic signaling in a ligand independent fashion. Understanding how integrin ?1?1 negatively controls T?RII pro-fibrotic action will contribute to the identification of 1) a novel crosstalk between integrin ?1?1 and T?RII; 2) a novel mechanism whereby T?RIImediated pro-fibrotic signaling can be negatively modulated; and 3) areas where integrin and/or T?RII functions can be specifically targeted to halt and ideally prevent CKD.
|Albertolle, Matthew E; Phan, Thanh T N; Pozzi, Ambra et al. (2018) Sulfenylation of Human Liver and Kidney Microsomal Cytochromes P450 and Other Drug-Metabolizing Enzymes as a Response to Redox Alteration. Mol Cell Proteomics 17:889-900|
|Sausville, Lindsay N; Gangadhariah, Mahesha H; Chiusa, Manuel et al. (2018) The Cytochrome P450 Slow Metabolizers CYP2C9*2 and CYP2C9*3 Directly Regulate Tumorigenesis via Reduced Epoxyeicosatrienoic Acid Production. Cancer Res 78:4865-4877|
|Brown, Kyle L; Banerjee, Surajit; Feigley, Andrew et al. (2018) Salt-bridge modulates differential calcium-mediated ligand binding to integrin ?1- and ?2-I domains. Sci Rep 8:2916|
|Borza, Corina M; Pozzi, Ambra; Plosa, Erin J (2018) Discoidin Domain Receptor 2, a Potential Therapeutic Target in Lung Fibrosis. Am J Respir Cell Mol Biol 59:277-278|
|Sausville, Lindsay N; Jones, Carissa C; Aldrich, Melinda C et al. (2017) Genetic variation in the eicosanoid pathway is associated with non-small-cell lung cancer (NSCLC) survival. PLoS One 12:e0180471|
|Albertolle, Matthew E; Kim, Donghak; Nagy, Leslie D et al. (2017) Heme-thiolate sulfenylation of human cytochrome P450 4A11 functions as a redox switch for catalytic inhibition. J Biol Chem 292:11230-11242|
|Borza, Corina M; Su, Yan; Tran, Truc-Linh et al. (2017) Discoidin domain receptor 1 kinase activity is required for regulating collagen IV synthesis. Matrix Biol 57-58:258-271|
|Kothiwale, Sandeepkumar; Borza, Corina; Pozzi, Ambra et al. (2017) Quantitative Structure-Activity Relationship Modeling of Kinase Selectivity Profiles. Molecules 22:|
|Mathew, Sijo; Palamuttam, Riya J; Mernaugh, Glenda et al. (2017) Talin regulates integrin ?1-dependent and -independent cell functions in ureteric bud development. Development 144:4148-4158|
|Williams, Ashley S; Trefts, Elijah; Lantier, Louise et al. (2017) Integrin-Linked Kinase Is Necessary for the Development of Diet-Induced Hepatic Insulin Resistance. Diabetes 66:325-334|
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