TGFβ family members, including TGFβs and BMPs, play a key role during embryonic development, in processes of cell differentiation[2]. RepSox is a selective inhibitor of TGF-β type I receptor (also called as TβRI or ALK5) with IC50 value of 4nM (measured by ALK5 autophosphorylation assay)[1]. RepSox is mainly used in cell reprogramming and differentiation as following: 1. direct reprogramming of mouse fibroblasts into cardiomyocytes, combined with CHIR99021, Forskolin, VPA, Parnate, TTNPB and DZnep[3]. 2. replaces Sox2 in reprogramming by inducing Nanog[4].
作用机制
RepSox can bound to the ATP binding site of the kinase domain of ALK5.[1]
RepSox 细胞实验
Cell Line
Concentration
Treated Time
Description
References
Mouse embryonic fibroblasts (MEFs)
1 µM
10 days
Induction of compact cell colonies and activation of Sox2 expression
Cell Res. 2014 Jun;24(6):665-79.
Mouse tail-tip fibroblasts (TTFs)
1 µM
10 days
Upregulation of Sox2 expression and formation of neural progenitor-like cells
Cell Res. 2014 Jun;24(6):665-79.
Mouse embryonic fibroblasts (MEFs)
7.5 µM
14-17 days
RepSox, as a TGF-β inhibitor, in combination with p53DD and hRasG12V, significantly increased the efficiency of induced motor neuron (iMN) reprogramming, reduced genomic stress, and expanded the population of hypertranscribing and hyperproliferating cells (HHCs).
Cell Stem Cell. 2019 Oct 3;25(4):486-500.e9.
Human urinary cells (HUCs)
1 µM
20 days
Formation of compact cell colonies and induction of neural progenitor-like cells
Cell Res. 2014 Jun;24(6):665-79.
HCMEC/d3 cells
100 µM
24 hours
RepSox significantly increased Cldn5 mRNA expression and enhanced barrier properties, as indicated by increased transendothelial electrical resistance (TEER) and reduced permeability to 40 kDa FITC-dextran (FD40).
Proc Natl Acad Sci U S A. 2020 Aug 18;117(33):19854-19865.
Cardiomyocytes
20 µM
5 days
Evaluate the effects of RepSox on cardiomyocyte viability and proliferation, results showed that RepSox significantly improved cardiomyocyte survival and proliferation.
Adv Funct Mater. 2020 Oct 22;30(43):2004307.
Cardiac fibroblasts
20 µM
5 days
Evaluate the effects of RepSox on cardiac fibroblast proliferation and myodifferentiation, results showed that RepSox inhibited fibroblast proliferation and myodifferentiation.
Adv Funct Mater. 2020 Oct 22;30(43):2004307.
Endothelial cells
20 µM
5 days
Evaluate the effects of RepSox on endothelial cell proliferation and vascular network formation, results showed that RepSox significantly enhanced endothelial cell proliferation and vascular network formation.
Adv Funct Mater. 2020 Oct 22;30(43):2004307.
Human epidermal stem cells (hESC)
5 µM
6 to 8 days
Conversion of human epidermal stem cells into intestinal goblet cells with an efficiency of ~95%
RepSox replaces Sox2 in reprogramming by inhibiting Tgf-β signaling and inducing Nanog expression.
Cell Stem Cell. 2009 Nov 6;5(5):491-503.
SH-SY5Y/6TR(EU)/pTrex-Dest-30/MYCN (SY5Y-MYCN)
1 - 100 nM
72 hours
RepSox significantly attenuated RA-mediated neuronal differentiation, indicating that TGF-β signaling inhibition plays a crucial role in retinoid-mediated differentiation in neuroblastoma cells.
Genome Med. 2017 Feb 10;9(1):15.
Adult tail tip fibroblasts
25 µM
RepSox effectively replaces Sox2 in adult tail tip fibroblasts, inducing reprogramming.
Cell Stem Cell. 2009 Nov 6;5(5):491-503.
RepSox 动物实验
Species
Animal Model
Administration
Dosage
Frequency
Description
References
C57BL/6J mice
Kainic acid-induced epilepsy model via intrahippocampal injection
Intraperitoneal injection
10 mg/kg
Every other day for 7 days
RepSox significantly reduced the severity of seizures and attenuated blood-brain barrier disruption.
Nat Commun. 2022 Apr 14;13(1):2003.
Rats
Myocardial infarction model
Intramyocardial injection
20 mg/mL
Single injection, lasting 5 weeks
Evaluate the therapeutic effects of RepSox on myocardial infarction in rats, results showed that RepSox significantly improved cardiac function, reduced fibrosis and inflammatory response, and promoted angiogenesis.
Adv Funct Mater. 2020 Oct 22;30(43):2004307.
Mice
Neonatal mouse retina model
Exogenous delivery
3 mg/kg
Not specified
RepSox significantly altered vascular patterning in the neonatal mouse retina, reducing the propagation of the angiogenic front but not changing the number of tip-cell microvilli.
Proc Natl Acad Sci U S A. 2020 Aug 18;117(33):19854-19865.
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