(124) A Novel JNK Inhibitor Reduces Inflammation and Improves Decidualization in Primary-Derived 2D and 3D Models of Endometriosis

Genesis Herrera^1,2, Tirupataiah Sirupangi^1,2, Chandrashekhar Madasu^1,2, Fei Yuan^1,2, Suni Tang^1,2, Caterina Clementi⁴, Kiran L. Sharma^1,2, Kurt M. Bohren^1,2, Linda Alpuing Radilla³, Xiaoming Guan³, Martin M. Matzuk^1,2, Stephen Palmer^1,2,4, Diana Monsivais^1,2

¹Department of Pathology & Immunology; ²Center for Drug Discovery; ³Obstetrics and Gynecology, Baylor College of Medicine, Houston, TX, 77030, USA.

⁴Celmatix, Inc. USA.

Abstract Text:

There is a medical need to develop non-hormonal purpose-designed therapeutic options for endometriosis that are safe, effective, and do not interfere with fertility. Because of its crucial pro-inflammatory role, the Jun N-terminal kinases (JNK1, -2, -3) are a clinically validated target for endometriosis. Novel JNK inhibitors, CDD-2728 and CDD-3013, were characterized by our center to be highly specific, cell-permeable, well-tolerated and effective in allograft disease models. We hypothesize that treating endometriosis with these JNK inhibitors will provide unprecedented treatment of the pain and infertility associated with the disease.

Previously reported 2D- and 3D- primary and immortalized cellular models of endometriosis were used to reveal the potency of CDD-2728 and CDD-3013 to suppress cytokine-induced inflammation while permitting progesterone-dependent decidualization. In vitro decidualization was induced with estradiol, medroxyprogesterone acetate, and cyclic AMP (EPC). Bulk RNA sequencing and quantitative RT-PCR were used in peritoneal endometriotic epithelial cells (12Z), primary eutopic stromal cells, endometriotic stromal cells, and 3D epithelial endometrial organoids from endometriosis patients.

Inhibition of JNK with CDD-2728 or CDD-3013 suppressed pro-inflammatory signaling pathways induced by IL-1b in various in vitro endometriosis models. Treatment of 12Z cells with IL-1b established an expression profile of inflammation (3405 genes) compared to vehicle ( >1.2, < 0.8, FDR < 0.05). When compared to IL-1b-treated 12Z cells, 1µM CDD-2728 changed the expression of 333 genes while 1µM CDD-3013 impacted 3452 genes ( >1.2, < 0.8, FDR < 0.05). Specifically, CDD-2728 or CDD-3013 suppressed IL-1b-induced pro-inflammatory genes, MMP3, IL8, TNC, and canonical (DUSP1,-3, -8) and non-canonical JNK genes (FGF5, KLF5). Similarly, 1µM CDD-2728 and CDD-3013, suppressed pro-inflammatory signaling pathways, “TNF signaling,” and “NF Kappa b,” in 3D endometrial epithelial organoids from endometriosis patients. When added to EPC-treated eutopic stromal cells from endometriosis patients, 1µM CDD-2728 and CDD-3013 increased PRL, IGFBP1, and FOXO1, relative to EPC treatment alone.

JNK inhibitors, CDD-2728 and CDD-3013, suppress pro-inflammatory signaling. JNK inhibition does not impair decidualization and increases progesterone-dependent decidual markers in primary stromal cells from endometriosis patients.

Funding from NICHD HD099341, HD105800, and Burroughs Wellcome Fund.