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Updated: May 5, 2026
Suppression of Pro-fibrotic Signaling Potentiates Factor-mediated Reprogramming of Mouse Embryonic Fibroblasts into Induced Cardiomyocytes
Published on: June 3, 2018
Single-target RNA interference for the blockade of multiple interacting proinflammatory and profibrotic pathways in
Juliane Tank1, Diana Lindner1, Xiaomin Wang1
1Dept. of Cardiology and Pneumology, CharitéCentrum11 (Cardiovascular Medicine), CBF, Germany.
Abstract:
Therapeutic targets of broad relevance are likely located in pathogenic pathways common to disorders of various etiologies. Screening for targets of this type revealed CCN genes to be consistently upregulated in multiple cardiomyopathies. We developed RNA interference (RNAi) to silence CCN2 and found this single-target approach to block multiple proinflammatory and profibrotic pathways in activated primary cardiac fibroblasts (PCFBs). The RNAi-strategy was developed in murine PCFBs and then investigated in "individual" human PCFBs grown from human endomyocardial biopsies (EMBs). Screening of short hairpin RNA (shRNA) sequences for high silencing efficacy and specificity yielded RNAi adenovectors silencing CCN2 in murine or human PCFBs, respectively. Comparison of RNAi with CCN2-modulating microRNA (miR) vectors expressing miR-30c or miR-133b showed higher efficacy of RNAi. In murine PCFBs, CCN2 silencing resulted in strongly reduced expression of stretch-induced chemokines (Ccl2, Ccl7, Ccl8), matrix metalloproteinases (MMP2, MMP9), extracellular matrix (Col3a1), and a cell-to-cell contact protein (Cx43), suggesting multiple signal pathways to be linked to CCN2. Immune cell chemotaxis towards CCN2-depleted PCFBs was significantly reduced. We demonstrate here that this RNAi strategy is technically applicable to "individual" human PCFBs, too, but that these display individually strikingly different responses to CCN2 depletion. Either genomically encoded factors or stable epigenetic modification may explain different responses between individual PCFBs. The new RNAi approach addresses a key regulator protein induced in cardiomyopathies. Investigation of this and other molecular therapies in individual human PCBFs may help to dissect differential pathogenic processes between otherwise similar disease entities and individuals.
Insights
Researchers developed RNA interference (RNAi) to silence CCN2, a key gene in cardiomyopathies. This approach effectively reduced pro-inflammatory and pro-fibrotic pathways in cardiac cells, offering a potential therapeutic strategy for heart conditions.
Area of Science:
- Cardiovascular Biology
- Molecular Medicine
- Gene Therapy
Background:
- Therapeutic targets relevant across diverse etiologies are often found in common pathogenic pathways.
- CCN genes, particularly CCN2, are consistently upregulated in various cardiomyopathies.
Purpose of the Study:
- To investigate CCN2 as a therapeutic target for cardiomyopathies using RNA interference (RNAi).
- To assess the efficacy of CCN2 silencing in blocking pro-inflammatory and pro-fibrotic pathways in cardiac fibroblasts.
Main Methods:
- Developed RNA interference (RNAi) strategies using short hairpin RNA (shRNA) adenovectors to silence CCN2.
- Tested RNAi efficacy in murine and human primary cardiac fibroblasts (PCFBs) derived from endomyocardial biopsies (EMBs).
- Compared RNAi efficacy with microRNA (miR)-based vectors (miR-30c, miR-133b).
Main Results:
- CCN2 silencing via RNAi effectively blocked multiple pro-inflammatory and pro-fibrotic pathways in PCFBs.
- Reduced expression of chemokines, matrix metalloproteinases, extracellular matrix components, and Cx43 was observed.
- Immune cell chemotaxis towards CCN2-depleted PCFBs was significantly reduced.
- RNAi strategy demonstrated applicability to individual human PCFBs, revealing differential responses.
Conclusions:
- CCN2 is a key regulator protein induced in cardiomyopathies, targeted effectively by RNAi.
- RNAi-mediated CCN2 silencing presents a promising single-target therapeutic approach for cardiomyopathies.
- Investigating therapies in individual human PCFBs is crucial for understanding differential pathogenic processes.
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