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Mitochondrial transfer mediates endothelial cell engraftment through…
Mitochondrial transfer mediates endothelial cell engraftment through mitophagy
introduction
Ischaemic diseases
basic
critical limb ischaemia
necessitate surgical interventions
vascular grafts to revascularize tissues distal to blockages
myocardial infarction
question
inadequate revascularization persists
delivery of angiogenic growth factors to stimulate local angiogenesis has been investigated
limited by the disrupted tissue microenvironment in ischaemic regions
EC therapies
key challenge ： the requirement for a secondary cell type to support engraftment
ECs co-transplantation with perivascular cells is essential for robust engraftment and functional blood vessel formation in vivo
Sources of perivascular cells include smooth muscle cells, pericytes, fibroblasts and MSCs
the use of multiple cell types complicates clinical translation and increases the complexity of phase I studies, and thus there is a reluctance to use two or more different cell types in clinical trials.
perivascular cells
perivascular cells facilitate EC engraftment are not fully understood
MSCs transfer mitochondria to ECs through tunnelling nanotubes (TNTs) during transplantation
aligns with previous findings of the transfer of mitochondria from MSCs to other cells
novel strategy
pre-emptively transplanting exogenous mitochondria into ECs to enhance engraftment in vivo
induced a transient cytoprotective effect through mitophagy
nabling the engraftment of ECs without the support of MSCs
Hypothesis
Q1: Whether EC engraftment requires stromal cells
Q2: Whether mitochondrial transfer of MSC facilitate EC transplantation
Q3: Whether exogenous mitochondrial transfer facilitate EC transplantation
Q4: the reason that mitochondrial transfer promotes EC transplantation
Results
A1
EC engraftment requires stromal cells
xenograft model
microvessel density
lucif-ECs
MSCs secrete a range of angiogenic factors
microvessel density
flow cytometry at 24 h
H&E staining
Proteomic dot blotting
A2
Mitochondrial transfer from MSCs to ECs via TNTs
MSCs were transfected with a piggyBac transposon containing a mito-tag
mitochondria DsRed+
Histological examination
Flow cytometric
immunofluorescence analyses
grafts seeded with ECs and mitoRed-MSCs
TNT formation was largely unidirectional
Mitochondrial transfer enables EC engraftment
shRNA silencing of TNFAIP2
silencing the expression of mitochondrial Rho GTPase 1 (MIRO1; also known as RHOT1)
A3
Artificial mitochondrial transplantation into ECs
Flow cytometry analysis
Transmission electron microscopy (TEM)
metabolic activities
resistance to apoptosis
increased cellular mobility
A4
Exogenous mitochondria are depolarized
mtDNA-free mitochondria enhance EC function
Transplanted mitochondria trigger mitophagy
RNA-seq
qPCR
Mitophagy mediates EC engraftment
学习点：从结构到功能，同时添加反向检测
不足点，分子机制可以继续深入挖掘