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Abstract: SA-PO745

Reduction of Systemic Inflammation by Notch3 Inhibition in HIV-Associated Nephropathy

Session Information

Category: Glomerular Diseases

  • 1304 Glomerular Diseases: Podocyte Biology

Authors

  • Thornton, Mackenzie, University of Kansas Medical Center Department of Internal Medicine, Kansas City, Kansas, United States
  • Sommer, Nicole, University of Kansas Medical Center Department of Internal Medicine, Kansas City, Kansas, United States
  • Mcgonigle, Mercedes Bravo, University of Kansas Medical Center Department of Internal Medicine, Kansas City, Kansas, United States
  • Dinh Phan, Johnny, University of Kansas Medical Center Department of Internal Medicine, Kansas City, Kansas, United States
  • Wang, Tao, The University of Manchester, Manchester, Manchester, United Kingdom
  • Singhal, Pravin C., Northwell Health, New Hyde Park, New York, United States
  • Fields, Timothy A., University of Kansas Medical Center Department of Internal Medicine, Kansas City, Kansas, United States
  • Sharma, Madhulika, University of Kansas Medical Center Department of Internal Medicine, Kansas City, Kansas, United States
Background

Anti-retroviral therapy (ART) suppresses HIV-associated Nephropathy (HIVAN) but does not protect from continuous generation of viral proteins and inflammation. We need therapies that inhibit viral replication and inflammation simultaneously to avoid progression into chronic kidney disease (CKD). We have previously shown that pan Notch inhibition ameliorated disease progression in the Tg26 mouse model of HIVAN. Here, we determined specifically the role of Notch3 pathway in HIVAN pathogenesis.

Methods

We labelled renal sections from HIVAN patients and mouse model (Tg26) for Notch3 via immunohistochemistry. We determined if HIV-1 induces Notch3 activation. We generated Tg26 mice with global Notch3 knockout (Tg-N3KO) and characterized the kidneys, followed by RNA sequencing. Macrophage- podocyte interactions were then studied. Soluble TNF alpha levels were evaluated using ELISA.

Results

Notch3 was activated in glomerular, tubular and interstitial cells of HIVAN biopsies and Tg26 mice. Notch3 expression was induced when podocytes were transfected with HIV-1 construct. Genetic knockout of Notch3 (N3KO) in Tg26 (Tg-N3KO) mice resulted in a significant increase in the life span. This was associated with marked improvement in glomerular and tubular injury and renal function. A striking reduction in infiltrating immune cells was observed. RNA sequencing and validation data indicated a marked reduction of macrophage markers in Tg-N3KO mice verses Tg26 mice. We then isolated and cultured bone marrow derived macrophages from Tg26 and Tg-N3KO mice and found that Notch3 and Notch ligands, Jagged 1 and Delta like 4 (Dll4) were markedly upregulated in Tg26 mice. N3KO normalized them. Conditioned macrophage media from BMDM derived from Tg26 mice resulted in Notch activation of podocytes. Finally, Notch3 deletion not only affected the kidneys but reduced the systemic expression of soluble inflammatory factor tumor necrosis factor alpha (TNF-alpha).

Conclusion

Notch 3 deletion in HIVAN model reduced kidney injury, improved renal function, reduced macrophage infiltration and reduced systemic TNF alpha levels which may collectively be responsible for the increase life span of the Tg26 mice. Thus, inhibition of Notch3 may constitute an attractive therapeutic strategy in HIV related CKDs.

Funding

  • NIDDK Support