Synergistic hyperactivation of both mTORC1 and mTORC2 underlies the neural abnormalities of PTEN-deficient human neurons and cortical organoids

Navroop K Dhaliwal; Octavia Yifang Weng; Xiaoxue Dong; Afrin Bhattacharya; Mai Ahmed; Haruka Nishimura; Wendy W Y Choi; Aditi Aggarwal; Bryan W Luikart; Qiang Shu; Xuekun Li; Michael D Wilson; Jason Moffat; Lu-Yang Wang; Julien Muffat; Yun Li

doi:10.1016/j.celrep.2024.114173

Synergistic hyperactivation of both mTORC1 and mTORC2 underlies the neural abnormalities of PTEN-deficient human neurons and cortical organoids

Cell Rep. 2024 May 28;43(5):114173. doi: 10.1016/j.celrep.2024.114173. Epub 2024 May 2.

Authors

Navroop K Dhaliwal¹, Octavia Yifang Weng², Xiaoxue Dong³, Afrin Bhattacharya⁴, Mai Ahmed¹, Haruka Nishimura⁴, Wendy W Y Choi⁵, Aditi Aggarwal⁴, Bryan W Luikart⁶, Qiang Shu⁷, Xuekun Li⁸, Michael D Wilson⁵, Jason Moffat⁹, Lu-Yang Wang¹⁰, Julien Muffat¹¹, Yun Li¹²

Affiliations

¹ Program in Developmental and Stem Cell Biology, The Hospital for Sick Children, 686 Bay Street, Toronto, ON M5G 0A4, Canada.
² Program in Developmental and Stem Cell Biology, The Hospital for Sick Children, 686 Bay Street, Toronto, ON M5G 0A4, Canada; Program in Neurosciences and Mental Health, The Hospital for Sick Children, 686 Bay Street, Toronto, ON M5G 0A4, Canada; Department of Physiology, University of Toronto, 1 King's College Circle, Toronto, ON M5S 1A8, Canada.
³ Program in Developmental and Stem Cell Biology, The Hospital for Sick Children, 686 Bay Street, Toronto, ON M5G 0A4, Canada; The Children's Hospital, National Clinical Research Center for Child Health, School of Medicine, Zhejiang University, Hangzhou 310052, China; The Institute of Translational Medicine, School of Medicine, Zhejiang University, Hangzhou 310029, China.
⁴ Program in Developmental and Stem Cell Biology, The Hospital for Sick Children, 686 Bay Street, Toronto, ON M5G 0A4, Canada; Department of Molecular Genetics, University of Toronto, 1 King's College Circle, Toronto, ON M5S 1A8, Canada.
⁵ Program in Developmental and Stem Cell Biology, The Hospital for Sick Children, 686 Bay Street, Toronto, ON M5G 0A4, Canada; Department of Molecular Genetics, University of Toronto, 1 King's College Circle, Toronto, ON M5S 1A8, Canada; Program in Genetics and Genome Biology, The Hospital for Sick Children, 686 Bay Street, Toronto, ON M5G 0A4, Canada.
⁶ Department of Molecular and Systems Biology, Geisel School of Medicine, Dartmouth College, Hanover, NH 03755, USA.
⁷ The Children's Hospital, National Clinical Research Center for Child Health, School of Medicine, Zhejiang University, Hangzhou 310052, China.
⁸ The Children's Hospital, National Clinical Research Center for Child Health, School of Medicine, Zhejiang University, Hangzhou 310052, China; The Institute of Translational Medicine, School of Medicine, Zhejiang University, Hangzhou 310029, China.
⁹ Department of Molecular Genetics, University of Toronto, 1 King's College Circle, Toronto, ON M5S 1A8, Canada; Program in Genetics and Genome Biology, The Hospital for Sick Children, 686 Bay Street, Toronto, ON M5G 0A4, Canada.
¹⁰ Program in Neurosciences and Mental Health, The Hospital for Sick Children, 686 Bay Street, Toronto, ON M5G 0A4, Canada; Department of Physiology, University of Toronto, 1 King's College Circle, Toronto, ON M5S 1A8, Canada.
¹¹ Program in Neurosciences and Mental Health, The Hospital for Sick Children, 686 Bay Street, Toronto, ON M5G 0A4, Canada; Department of Molecular Genetics, University of Toronto, 1 King's College Circle, Toronto, ON M5S 1A8, Canada.
¹² Program in Developmental and Stem Cell Biology, The Hospital for Sick Children, 686 Bay Street, Toronto, ON M5G 0A4, Canada; Department of Molecular Genetics, University of Toronto, 1 King's College Circle, Toronto, ON M5S 1A8, Canada. Electronic address: yun.li@sickkids.ca.

PMID: 38700984
DOI: 10.1016/j.celrep.2024.114173

Abstract

Mutations in the phosphatase and tensin homolog (PTEN) gene are associated with severe neurodevelopmental disorders. Loss of PTEN leads to hyperactivation of the mechanistic target of rapamycin (mTOR), which functions in two distinct protein complexes, mTORC1 and mTORC2. The downstream signaling mechanisms that contribute to PTEN mutant phenotypes are not well delineated. Here, we show that pluripotent stem cell-derived PTEN mutant human neurons, neural precursors, and cortical organoids recapitulate disease-relevant phenotypes, including hypertrophy, electrical hyperactivity, enhanced proliferation, and structural overgrowth. PTEN loss leads to simultaneous hyperactivation of mTORC1 and mTORC2. We dissect the contribution of mTORC1 and mTORC2 by generating double mutants of PTEN and RPTOR or RICTOR, respectively. Our results reveal that the synergistic hyperactivation of both mTORC1 and mTORC2 is essential for the PTEN mutant human neural phenotypes. Together, our findings provide insights into the molecular mechanisms that underlie PTEN-related neural disorders and highlight novel therapeutic targets.

Keywords: CP: Cell biology; CP: Neuroscience; PTEN; autism; brain organoids; epilepsy; mTOR; neurodevelopmental disorders; neurons.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cell Proliferation
Cerebral Cortex / metabolism
Cerebral Cortex / pathology
Humans
Mechanistic Target of Rapamycin Complex 1* / metabolism
Mechanistic Target of Rapamycin Complex 2* / metabolism
Mutation / genetics
Neurons* / metabolism
Organoids* / metabolism
PTEN Phosphohydrolase* / genetics
PTEN Phosphohydrolase* / metabolism
Phenotype
Rapamycin-Insensitive Companion of mTOR Protein / genetics
Rapamycin-Insensitive Companion of mTOR Protein / metabolism
Regulatory-Associated Protein of mTOR / genetics
Regulatory-Associated Protein of mTOR / metabolism
Signal Transduction

Substances

PTEN Phosphohydrolase
Mechanistic Target of Rapamycin Complex 1
Mechanistic Target of Rapamycin Complex 2
PTEN protein, human
Rapamycin-Insensitive Companion of mTOR Protein
RICTOR protein, human
Regulatory-Associated Protein of mTOR
RPTOR protein, human