Hematopoietic Stem-Cell Senescence and Myocardial Repair: Coronary Artery Disease Genotype/Phenotype Analysis of Post-MI Myocardial Regeneration Response Induced by CABG/CD133+ Bone Marrow Hematopoietic Stem Cell Treatment in RCT PERFECT Phase 3
167 Pages Posted: 4 Jun 2020More...
Objective: Clonal hematopoiesis with mutational gene dysregulation of hematopoietic stem cell (HSC) precursors for immune or endothelial progenitor cell response is suspected to affect coronary artery disease bypass surgery (CABG) induced ischemic myocardial repair.
Design: Transcriptome and variant expression analysis was studied in the phase 3 PERFECT (post MI CABG and CD133 + BMSC or Placebo) trial in myocardial regeneration responders (R, n=14; ∆LVEF +16% day 180/0) and non-responders (NR, n=9; ∆LVEF -1∙1% day 180/0), then validated in independent patient cohort (n=14) as well as under mouse SH2B3/LNK antisense or knockout deficient conditions.
Findings: 1. Left ventricular functional recovery was accompanied by an increase in myocardial perfusion (R vs NR, p<0∙05). R differed from NR in a total of 161 genes in differential expression (n=23, q<0,05) and 872 genes in coexpression analysis (n=23, q<0,05). Machine Learning clustering analysis revealed distinct R vs NR preoperative gene-expression signature in peripheral blood. RvsNR ML-top selected genes were correlated to SH2B3 (p<0∙05). Mutation signature analysis performed by RNASeq revealed increased specific variants in NR vs R. (NR: 224 genes with 268 exon regions; R: 48 genes with 56 exon regions). 2. RvsNR ML-selected gene expression and phenotype was correlated to SH2B3 deficient mouse models. Competitive bone-marrow transplanted mice SH2B3/LNK silenced HSC clones displayed significant overgrowth of myeloid and immune cells in bone marrow, peripheral blood and tissue at day 160 after BMT. SH2B3/LNK-/- mice demonstrated enhanced cardiac repair through augmenting the kinetics of BM-derived EPCs, increased capillary density in ischemic myocardium and reduced left ventricular fibrosis with preserved cardiac function. 3. Validation analysis in 14 additional patients revealed 85% R vs NR prediction accuracy.
Interpretation: Myocardial repair is affected by HSC gene response. Somatic mutation in adaptor proteins may alter response by clonal overgrowth as shown for SH2B3/LNK. ML guided predictions can be utilized to identify and predict pathological HSC response.
Trial Registration: ClinicalTrials.gov NCT00950274
Funding Statement: German Ministry of Research and Education (BMBF): FKZ0312138A, FKZ031L0106C, EU ESF/IV-WM-B34-0011/08, ESF/IV-WM-B34-0030/10, and Miltenyi Biotec GmbH, Bergisch-Gladbach, Germany. Japanese Ministry of Health : Health and Labour Sciences Research Grant (H14-trans-001, H17-trans-002)
Declaration of Interests: All authors declare no competing interests.
Ethics Approval Statement: Approval by the Ethical committee, Rostock University Medical Center 2009; No. HV-2009-0012.
Keywords: Clonal hematopoiesis of indeterminate pathology; CHIP: coronary artery disease; cardiac regeneration capacity; randomized double-blinded phase 3 multicentre trial; myocardial repair response; CD133; CD34; hematopoietic stem cell; endothelial progenitor cell; EPC; SH2B3; LNK adaptor; Lnk; SH2B3/LNK-/
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