Elsevier

EBioMedicine

Volume 57, July 2020, 102862
EBioMedicine

Research paper
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

https://doi.org/10.1016/j.ebiom.2020.102862Get rights and content
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Abstract

Background

Bone marrow stem cell clonal dysfunction by somatic mutation is suspected to affect post-infarction myocardial regeneration after coronary bypass surgery (CABG).

Methods

Transcriptome and variant expression analysis was studied in the phase 3 PERFECT trial post myocardial infarction CABG and CD133+ bone marrow derived hematopoetic stem cells showing difference in left ventricular ejection fraction (∆LVEF) myocardial regeneration Responders (n=14; ∆LVEF +16% day 180/0) and Non-responders (n=9; ∆LVEF -1.1% day 180/0). Subsequently, the findings have been validated in an independent patient cohort (n=14) as well as in two preclinical mouse models investigating SH2B3/LNK antisense or knockout deficient conditions.

Findings

1. Clinical: 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 RvsNR preoperative gene-expression signatures in peripheral blood acorrelated to SH2B3 (p<0.05). Mutation analysis revealed increased specific variants in RvsNR. (R: 48 genes; NR: 224 genes). 2. Preclinical: SH2B3/LNK-silenced hematopoietic stem cell (HSC) clones displayed significant overgrowth of myeloid and immune cells in bone marrow, peripheral blood, and tissue at day 160 after competitive bone-marrow transplantation into mice. SH2B3/LNK−/− mice demonstrated enhanced cardiac repair through augmenting the kinetics of bone marrow-derived endothelial progenitor cells, increased capillary density in ischemic myocardium, and reduced left ventricular fibrosis with preserved cardiac function. 3. Validation: Evaluation analysis in 14 additional patients revealed 85% RvsNR (12/14 patients) prediction accuracy for the identified biomarker signature.

Interpretation

Myocardial repair is affected by HSC gene response and somatic mutation. Machine Learning can be utilized to identify and predict pathological HSC response.

Funding

German Ministry of Research and Education (BMBF): Reference and Translation Center for Cardiac Stem Cell Therapy - FKZ0312138A and FKZ031L0106C, German Ministry of Research and Education (BMBF): Collaborative research center - DFG:SFB738 and Center of Excellence - DFG:EC-REBIRTH), European Social Fonds: 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)

Trial registration

ClinicalTrials.gov NCT00950274

Keywords

Clonal hematopoiesis of indeterminate pathology
CHIP
SH2B3
Myocardial regeneration
Cardiac stem cell therapy
Angiogenesis induction
Post myocardial infarction heart failure
Coronary bypass surgery
CABG
Machine learning

Abbreviation

ANCOVA
Analysis of covariance
Ang-1
Angiopoeitin 1
AUR
Arch user repository
AUC
Area under curve
BM
Bone marrow
BMMNC
Bone marrow mononuclear cell
BMSC
Bone marrow stem cells
BrdU
Brome deoxyuridine
CABG
Coronary Artery Bypass Graft
CAP-EPC
Concentrated Ambient Particles – Endothelial Progenitor Cells
CD
Cluster of Differentiation
CEC
Circulating endothelial cells, CEC panel, CDs measured in PB
CFU
Colony-forming unit
CHIP
Clonal hematopoiesis of indeterminate potential
CI
Confidence interval
c-KIT/CD117
Stem Cell Factor Receptor c-KIT, CD117
CLARA
Clustering for Large Applications
CPC
cardiac progenitor cell
CSC
cardiac stem cell
DE
Differential gene expression
DNAseq
desoxyribonucleid acid sequencing
EGF
Epidermal growth factor
EGFR
Epidermal growth factor receptor
ELISA
Enzyme-Linked Immunosorbent Assay
EPC
Endothelial Progenitor Cells, EPC panel, CDs measured in PB
EPO
Erythropoietin
EPOR
Erythropoietin receptor
FACS
Fluorescence activated cell sorter
FDR
False discovery rate
GADPH
Glyceraldehyde 3 phosphate dehydrogenase
GATA4
Transcriptional activator that binds to the consensus sequence 5′-AGATAG-3’
GFP
Green fluorescent protein
GMP
Good Manufacturing Practice
GWAS
Genome wide association study
HR
Hazard ratio
HIF
Hypoxia-Inducible Factor, transcription factor
HSC
Hemopoeitic stem cell
hu
human
ICH GCP
Tripartite Guidelines Guideline for Good Clinical Practice
IGF-1
Insuline-like Growth Factor 1
IGFBP
Insuline-like growth factor binding proteine
IHG
Analysis performed in accordance with ISHAGE guidelines
IL
Interleukin
InDel
mutation insertion or deletion variant
KSL
mouse bone marrow stem cell subpopulation c-KIT+ Sca-I+ lin-
LAD
Left anterior descending coronary artery, RIVA
LAS
Longitudinal axis strain
LCRC
Loss of cardiac regeneration capacity
LNK
SH2B adapter protein 3 (lymphocyte adapter protein)
LVEDV
Left Ventricular End Diastolic Volume
LVEF
Left Ventricular Ejection Fraction
LVESD
Left Ventricular End Systolic Dimension
m
mouse
MI
myocardial infarction
ML
Machine learning
MNC
Mononuclear cells
MRI
Magnetic resonance imaging
6MWT
6-Minute Walk Test
NGS
Next Generation Sequencing
NR
non-responder
PB
Peripheral blood
PBMNC
mononuclear cells isolated from peripheral blood
PCR
Polymerase chain reaction
PDGF
Platelet derived growth factor
PDFR
Platelet derived growth factor receptor
PEI
Paul-Ehrlich Institute
PI3K
Phosphoinositide-3-Kinase
PBMNC
Peripheral blood mononuclear cell
PPMC
Pearson Product Moment Correlation
qPCR
Quantitative polymerase chain reaction
R
responder
RFI
Reactome functional interaction
RNASeq
Ribonucleid acid sequencing
ROC
Receiver operating characteristics
RT-PCR
Reverse transcriptase polymerase chain reaction
RWMS
Regional wall motion score
SDF-1
Stromal Cell-derived Factor 1
SH2B3
LNK [Src homology 2-B3 (SH2B3)] belongs to a family of SH2-containing proteins with important adaptor functions
SCF
Stem Cell Factor
SNP
Single nucleotide polymorphism, variant
STEMI
ST- segment Elevation Infarction
SUSAR
Suspected Unexpected Serious Adverse Reaction
TiCoNE
Time course network enrichment
TNF
Tumor Necrosis Factor
t-SNE
t-distributed stochastic neighbour embedding
VCA
Virus-Capsid-Antigen
VEGF
Vascular Endothelial Growth Factor
VEGFR
Vascular Endothelial Growth Factor Receptor
WT
wild type
WGCNA
Weighted gene coexpression network analysis

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