Hexokinase 2-driven glycolysis in pericytes activates their contractility leading to tumor blood vessel abnormalities

Qiong Meng,Liangping Su,Ping-Pui Wong,Xue Jiang,Chao Liu,Xinbao Zhao,Sangqing Wu,Ya-Ming Meng,Xiaoyi Qiu,Yitian Chen,Minghui Wang,Cheng Huang,Xiangzhan Kong

doi:10.1038/s41467-021-26259-y

Abstract

Defective pericyte-endothelial cell interaction in tumors leads to a chaotic, poorly organized and dysfunctional vasculature. However, the underlying mechanism behind this is poorly studied. Herein, we develop a method that combines magnetic beads and flow cytometry cell sorting to isolate pericytes from tumors and normal adjacent tissues from patients with non-small cell lung cancer (NSCLC) and hepatocellular carcinoma (HCC). Pericytes from tumors show defective blood vessel supporting functions when comparing to those obtained from normal tissues. Mechanistically, combined proteomics and metabolic flux analysis reveals elevated hexokinase 2(HK2)-driven glycolysis in tumor pericytes, which up-regulates their ROCK2-MLC2 mediated contractility leading to impaired blood vessel supporting function. Clinically, high percentage of HK2 positive pericytes in blood vessels correlates with poor patient overall survival in NSCLC and HCC. Administration of a HK2 inhibitor induces pericyte-MLC2 driven tumor vasculature remodeling leading to enhanced drug delivery and efficacy against tumor growth. Overall, these data suggest that glycolysis in tumor pericytes regulates their blood vessel supporting role.

Highlights

Defective pericyte-endothelial cell interaction in tumors leads to a chaotic, poorly organized and dysfunctional vasculature
Our findings show that treatment with hexokinase 2 (HK2) inhibitor or depletion of pericyte-HK2 by shRNA can enhance blood flow and perfusion leading to increased intratumoral drug delivery and efficacy against tumor growth, whilst decreasing tumor hypoxia without affecting blood vessel density and pericyte coverage (Fig. 7)
We showed an elevated HK2-driven glycolysis in TPC as compared with NPC, which contributed to its abnormal blood vessel supporting function by activating ROCK2-MLC2 mediated contractility

Summary

Result

Development of the microbead activated vascular cell sorting enrichment method coupled with FACS. TPC’s BV supporting function, in vitro tube formation assay was performed by co-culturing HUVEC with NSCLC/HCC-derived TPC pre-treated with either placebo or ROCK inhibitor GSK 429286A in matrigel, indicating that pre-treatment with ROCK inhibitor significantly enhanced total tube length, branch point, and number (Fig. 5l–o). To determine whether 3-BP or/and doxorubicin could affect tumor cells mediated pericyte’s glycolytic change, we examined the effect of conditioned medium harvested from cancer cells pre-treated with 3-BP or/and doxorubicin on pericyte-HK2 and ROCK2 expression Both RT-PCR and western blot analysis indicated that the conditioned medium of 3-BP or/and DOX pre-treated tumor cells could still up-regulate HK2 and ROCK2 expression in NSCLC/ HCC-derived NPC as compared to untreated NPC group These results indicate that targeting pericyte-HK2 expression/activity could be an effective treatment strategy for cancer patients

Discussion

Findings

Methods