VHL suppresses autophagy and tumor growth through PHD1-dependent Beclin1 hydroxylation

Zheng Wang; Meisi Yan; Leiguang Ye; Qimin Zhou; Yuran Duan; Hongfei Jiang; Lei Wang; Yuan Ouyang; Huahe Zhang; Yuli Shen; Guimei Ji; Xiaohan Chen; Qi Tian; Liwei Xiao; Qingang Wu; Ying Meng; Guijun Liu; Leina Ma; Bo Lei; Zhimin Lu; Daqian Xu

doi:10.1038/s44318-024-00051-2

VHL suppresses autophagy and tumor growth through PHD1-dependent Beclin1 hydroxylation

EMBO J. 2024 Mar;43(6):931-955. doi: 10.1038/s44318-024-00051-2. Epub 2024 Feb 15.

Authors

Zheng Wang^#^{1

2}, Meisi Yan^#³, Leiguang Ye^#⁴, Qimin Zhou^#⁵, Yuran Duan^{1

2}, Hongfei Jiang⁶, Lei Wang^{1

2}, Yuan Ouyang^{7

8

9

10}, Huahe Zhang^{11

12}, Yuli Shen^{1

2}, Guimei Ji^{1

2}, Xiaohan Chen¹³, Qi Tian^{1

2}, Liwei Xiao^{1

2}, Qingang Wu^{1

2}, Ying Meng^{1

2}, Guijun Liu^{1

2}, Leina Ma⁶, Bo Lei^{14

15}, Zhimin Lu^{16

17}, Daqian Xu^{18

19}

Affiliations

¹ Zhejiang Provincial Key Laboratory of Pancreatic Disease, The First Affiliated Hospital, and Institute of Translational Medicine, Zhejiang University School of Medicine, Zhejiang University, 310029, Hangzhou, China.
² Cancer Center, Zhejiang University, 310029, Hangzhou, Zhejiang, China.
³ Department of Pathology, Harbin Medical University, Harbin, China.
⁴ Department of Medical Oncology, Harbin Medical University Cancer Hospital, Harbin, China.
⁵ Department of Plastic and Reconstructive Surgery, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, 200011, Shanghai, China.
⁶ Department of Oncology, Cancer Institute, The Affiliated Hospital of Qingdao University, Qingdao University, Qingdao Cancer Institute, 266061, Qingdao, Shandong, China.
⁷ Laboratory of Oral Microbiota and Systemic Diseases, Shanghai Ninth People's Hospital, College of Stomatology, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
⁸ National Center for Stomatology, Shanghai, China.
⁹ National Clinical Research Center for Oral Diseases, Shanghai, China.
¹⁰ Shanghai Key Laboratory of Stomatology, Shanghai, China.
¹¹ Department of Breast Surgery, Harbin Medical University Cancer Hospital, Harbin, China.
¹² NHC Key Laboratory of Cell Transplantation, The First Affiliated Hospital of Harbin Medical University, 150001, Harbin, Heilongjiang Province, China.
¹³ Department of Oncology, Harbin Medical University Cancer Hospital, Harbin Medical University, 150001, Harbin, Heilongjiang Province, China.
¹⁴ Department of Breast Surgery, Harbin Medical University Cancer Hospital, Harbin, China. leibo@hrbmu.edu.cn.
¹⁵ NHC Key Laboratory of Cell Transplantation, The First Affiliated Hospital of Harbin Medical University, 150001, Harbin, Heilongjiang Province, China. leibo@hrbmu.edu.cn.
¹⁶ Zhejiang Provincial Key Laboratory of Pancreatic Disease, The First Affiliated Hospital, and Institute of Translational Medicine, Zhejiang University School of Medicine, Zhejiang University, 310029, Hangzhou, China. zhiminlu@zju.edu.cn.
¹⁷ Cancer Center, Zhejiang University, 310029, Hangzhou, Zhejiang, China. zhiminlu@zju.edu.cn.
¹⁸ Zhejiang Provincial Key Laboratory of Pancreatic Disease, The First Affiliated Hospital, and Institute of Translational Medicine, Zhejiang University School of Medicine, Zhejiang University, 310029, Hangzhou, China. xudaqian@zju.edu.cn.
¹⁹ Cancer Center, Zhejiang University, 310029, Hangzhou, Zhejiang, China. xudaqian@zju.edu.cn.

^# Contributed equally.

Abstract

The Von Hippel-Lindau (VHL) protein, which is frequently mutated in clear-cell renal cell carcinoma (ccRCC), is a master regulator of hypoxia-inducible factor (HIF) that is involved in oxidative stresses. However, whether VHL possesses HIF-independent tumor-suppressing activity remains largely unclear. Here, we demonstrate that VHL suppresses nutrient stress-induced autophagy, and its deficiency in sporadic ccRCC specimens is linked to substantially elevated levels of autophagy and correlates with poorer patient prognosis. Mechanistically, VHL directly binds to the autophagy regulator Beclin1, after its PHD1-mediated hydroxylation on Pro54. This binding inhibits the association of Beclin1-VPS34 complexes with ATG14L, thereby inhibiting autophagy initiation in response to nutrient deficiency. Expression of non-hydroxylatable Beclin1 P54A abrogates VHL-mediated autophagy inhibition and significantly reduces the tumor-suppressing effect of VHL. In addition, Beclin1 P54-OH levels are inversely correlated with autophagy levels in wild-type VHL-expressing human ccRCC specimens, and with poor patient prognosis. Furthermore, combined treatment of VHL-deficient mouse tumors with autophagy inhibitors and HIF2α inhibitors suppresses tumor growth. These findings reveal an unexpected mechanism by which VHL suppresses tumor growth, and suggest a potential treatment for ccRCC through combined inhibition of both autophagy and HIF2α.

Keywords: Autophagy; Beclin1; Hydroxylation; VHL; ccRCC.

MeSH terms

Animals
Autophagy
Beclin-1* / genetics
Beclin-1* / metabolism
Carcinoma, Renal Cell* / metabolism
Cell Line, Tumor
Gene Expression Regulation, Neoplastic
Humans
Hydroxylation
Kidney Neoplasms* / metabolism
Mice
Procollagen-Proline Dioxygenase / metabolism
Von Hippel-Lindau Tumor Suppressor Protein* / genetics
Von Hippel-Lindau Tumor Suppressor Protein* / metabolism

Substances

Beclin-1
PHD1 protein, mouse
Procollagen-Proline Dioxygenase
VHL protein, human
Von Hippel-Lindau Tumor Suppressor Protein
EGLN2 protein, human
BECN1 protein, human

Abstract

MeSH terms

Substances

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