Details

Autor(en) / Beteiligte

• Sano, Toshikazu
• Ito, Tatsuo
• Ishigami, Shuta
• Bandaru, Srinivas
• Sano, Shunji

Titel

Intrinsic activation of cardiosphere-derived cells enhances myocardial repair

Ist Teil von

• The Journal of thoracic and cardiovascular surgery, 2022-04, Vol.163 (4), p.1479-1490.e5

Ort / Verlag

United States: Elsevier Inc

Erscheinungsjahr

2022

Quelle

Free E-Journal (出版社公開部分のみ）

Beschreibungen/Notizen

• Permanent loss of cardiomyocytes after myocardial infarction results in irreversible damage to cardiac function. The present study aims to enhance the cardiomyogenic efficiency of cardiosphere-derived cells (CDCs) to develop into large populations of cardiomyocytes by intrinsic activation of cardio-specific differentiation factors (Gata4, Mef2c, Nkx2-5, Hand2, and Tnnt2) by a CRISPR/dCas9 assisted transcriptional enhancement system. Exhaustive screening was performed to identify the specific sequences in endogenous regulatory regions (enhancers and promoters) responsible for transcriptional activation of the target genes, via a CRISPR/dCas9 system fused with transcriptional activator VP64 (CRISPR-dCas9-VP64). In a rat model of acute myocardial infarction, we compared the regenerative potential and functional benefits of CDCs with or without transcriptional activation. We identified a panel of specific CRISPR RNA targeting the enhancers and promoters, which demonstrated significantly higher expression of differentiation factors of Gata4, Hand2, and Tnnt2. The group of CDCs with transcriptional activator VP64 (CDC with VP64) showed significant improvement in the left ventricular ejection fraction (61.9% vs 52.5% and 44.1% in the CDC without transcriptional activation group and control) and decreased scar area in the heart. We have identified endogenous regulatory regions responsible for an intrinsic activation of cardio-specific differentiation factors assisted via a CRISPR/dCas9 gene transcriptional system. The CRISPR/dCas9 system may provide an efficient and effective means of regulating Tnnt2 gene activation within stem cells. Subsequently, this system can be used to enhance transplanted CDCs differentiation potential within ischemic myocardia to better therapeutic outcomes of patients with ischemic heart disease. Screening was performed to identify the specific sequences responsible for transcriptional activation of the target genes via a CRISPR/dCas9 system fused with transcriptional activator VP64 (CRISPR-dCas9-VP64). We compared the regenerative potential and functional benefits of cardiosphere-derived cells (CDCs) with or without transcriptional activation in a rat model of acute myocardial infarction. The group of CDCs with transcriptional activator VP64 (CDC with VP64) showed significant improvement in the left ventricular ejection fraction and decreased scar area in the heart. PBS, Phosphate buffered saline. [Display omitted]