Abstract Body (Do not enter title and authors here): Background: Cardiac fibrosis is a critical and independent driver of heart failure (HF), characterized by excessive extracellular matrix (ECM) deposition. Decreased CDCP1 (CUB domain-containing protein 1) expression has been associated with myocardial recovery in human genome wide association studies by attenuating PDGF-BB driven cardiac fibroblast proliferation with no effect on canonical TGF-beta mediated transdifferentiation in-vitro. In-vivo, Cdcp1 deletion significantly reduced cardiac fibrosis and dysfunction in an angiotensin II/phenylephrine-induced pressure overload mouse model. However, its upstream regulators and its downstream effects on molecular signaling pathways that modulate ECM remodeling are unknown.
Hypothesis: CDCP1 promotes human cardiac fibroblast mediated ECM remodeling in the HF microenvironment.
Methods: Human ventricular fibroblasts (HVF) were treated with PDGF-BB (20 ng/ml), TGF-β (10 ng/ml), or Angiotensin II (100 ng/ml) for 48 hours. CDCP1 knockdown (KD) was achieved through siRNA transfection. Gene expression analysis was performed using qRT-PCR and protein quantification by Western blot. Both intact HVF and decellularized ECM were immunostained for collagen 1 to evaluate for ECM compositional changes.
Results: Individual and combination treatments with fibrotic stimuli significantly increased CDCP1 expression (Figure 1A-1B). CDCP1 KD resulted in reduction of key ECM markers COL1A1, CTGF, and LOX (Figure 1C) across all fibrotic stimuli. Immunostaining of HVF after treatment with fibrotic stimuli showed reduced collagen 1 production with CDCP1 KD (Figure 1D). Western blot analysis showed increased phosphorylation of Src (tyr416) with 6h of treatment with fibrotic stimuli which was nullified by silencing of CDCP1 (Figure 1E).
Conclusion: Our findings demonstrate that CDCP1 is necessary for ECM synthesis and Src signaling across multiple profibrotic pathways. Selective targeting of CDCP1 to modulate Src signaling pathways could provide novel therapeutic avenues for HF and fibrosis treatment.