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Published on: July 22, 2020
Targeting Src SH3 domain-mediated glycolysis of HSC suppresses transcriptome, myofibroblastic activation, and
Yuanguo Wang1, Xianghu Wang1,2, Bing Bai1
1Tumor Microenvironment and Metastasis, the Hormel Institute, University of Minnesota, Austin, Minnesota, USA.
Background And Aims:
Transforming growth factor-beta 1 (TGFβ1) induces HSC activation into metastasis-promoting cancer-associated fibroblasts (CAFs), but how the process is fueled remains incompletely understood. We studied metabolic reprogramming induced by TGFβ1 in HSCs.
Approaches And Results:
Activation of cultured primary human HSCs was assessed by the expression of myofibroblast markers. Glucose transporter 1 (Glut1) of murine HSC was disrupted by Cre recombinase/LoxP sequence derived from bacteriophage P1 recombination (Cre/LoxP). Plasma membrane (PM) Glut1 and glycolysis were studied by biotinylation assay and the Angilent Seahorse XFe96 Analyzer. S.c. HSC/tumor co-implantation and portal vein injection of MC38 colorectal cancer cells into HSC-specific Glut1 knockout mice were performed to determine in vivo relevance. Transcriptome was obtained by RNA sequencing of HSCs and spatialomics with MC38 liver metastases. TGFβ1-induced CAF activation of HSCs was accompanied by elevation of PM Glut1, glucose uptake, and glycolysis. Targeting Glut1 or Src by short hairpin RNA, pharmacologic inhibition, or a Src SH3 domain deletion mutant abrogated TGFβ1-stimulated PM accumulation of Glut1, glycolysis, and CAF activation. Mechanistically, binding of the Src SH3 domain to SH3 domain-binding protein 5 led to a Src/SH3 domain-binding protein 5/Rab11/Glut1 complex that activated Rab11-dependent Glut1 PM transport under TGFβ1 stimulation. Deleting the Src SH3 domain or targeting Glut1 of HSCs by short hairpin RNA or Cre recombinase/LoxP sequence derived from bacteriophage P1 recombination suppressed CAF activation in mice and MC38 colorectal liver metastasis. Multi-omics revealed that Glut1 deficiency in HSCs/CAFs suppressed HSC expression of tumor-promoting factors and altered MC38 transcriptome, contributing to reduced MC38 liver metastases.
Conclusion:
The Src SH3 domain-facilitated metabolic reprogramming induced by TGFβ1 represents a target to inhibit CAF activation and the pro-metastatic liver microenvironment.
Insights
Transforming growth factor-beta 1 (TGFβ1) drives hepatic stellate cell (HSC) activation into cancer-associated fibroblasts (CAFs). Targeting glucose transporter 1 (Glut1) and Src signaling inhibits this metabolic reprogramming, suppressing liver metastasis.
Area of Science:
- Cell Biology
- Metabolism
- Cancer Research
Background:
- Transforming growth factor-beta 1 (TGFβ1) promotes hepatic stellate cell (HSC) activation into cancer-associated fibroblasts (CAFs).
- The metabolic reprogramming fueling this HSC activation remains poorly understood.
- This study investigates the role of glucose metabolism in TGFβ1-induced HSC activation.
Purpose of the Study:
- To elucidate the mechanisms of metabolic reprogramming in HSCs induced by TGFβ1.
- To determine the role of glucose transporter 1 (Glut1) and Src signaling in CAF activation.
- To evaluate the therapeutic potential of targeting these pathways in liver metastasis.
Main Methods:
- Assessed HSC activation markers and measured glucose uptake and glycolysis.
- Utilized genetic manipulation (Cre/LoxP) to disrupt Glut1 in HSCs.
- Employed in vivo models including HSC/tumor co-implantation and orthotopic injections in knockout mice.
- Performed RNA sequencing and spatialomics on HSCs and liver metastases.
Main Results:
- TGFβ1 increased plasma membrane Glut1, glucose uptake, and glycolysis in HSCs, driving CAF activation.
- Targeting Glut1 or Src signaling abrogated TGFβ1-induced CAF activation.
- A Src/SH3 domain-binding protein 5/Rab11/Glut1 complex mediates Glut1 transport to the plasma membrane.
- HSC-specific Glut1 deletion suppressed CAF activation and reduced liver metastasis in vivo.
- Multi-omics analysis revealed that Glut1 deficiency altered HSC and tumor cell transcriptomes, reducing metastasis.
Conclusions:
- Src SH3 domain-mediated metabolic reprogramming is crucial for TGFβ1-induced CAF activation.
- Targeting the Src/Glut1 axis offers a potential strategy to inhibit CAF activation and liver metastasis.
- Inhibition of HSC metabolic reprogramming can disrupt the pro-metastatic liver microenvironment.
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