Rapid iPSC inclusionopathy models shed light on formation, consequence, and molecular subtype of α-synuclein inclusions

Isabel Lam; Alain Ndayisaba; Amanda J Lewis; YuHong Fu; Giselle T Sagredo; Anastasia Kuzkina; Ludovica Zaccagnini; Meral Celikag; Jackson Sandoe; Ricardo L Sanz; Aazam Vahdatshoar; Timothy D Martin; Nader Morshed; Toru Ichihashi; Arati Tripathi; Nagendran Ramalingam; Charlotte Oettgen-Suazo; Theresa Bartels; Manel Boussouf; Max Schäbinger; Erinc Hallacli; Xin Jiang; Amrita Verma; Challana Tea; Zichen Wang; Hiroyuki Hakozaki; Xiao Yu; Kelly Hyles; Chansaem Park; Xinyuan Wang; Thorold W Theunissen; Haoyi Wang; Rudolf Jaenisch; Susan Lindquist; Beth Stevens; Nadia Stefanova; Gregor Wenning; Wilma D J van de Berg; Kelvin C Luk; Rosario Sanchez-Pernaute; Juan Carlos Gómez-Esteban; Daniel Felsky; Yasujiro Kiyota; Nidhi Sahni; S Stephen Yi; Chee Yeun Chung; Henning Stahlberg; Isidro Ferrer; Johannes Schöneberg; Stephen J Elledge; Ulf Dettmer; Glenda M Halliday; Tim Bartels; Vikram Khurana

doi:10.1016/j.neuron.2024.06.002

Rapid iPSC inclusionopathy models shed light on formation, consequence, and molecular subtype of α-synuclein inclusions

Neuron. 2024 Sep 4;112(17):2886-2909.e16. doi: 10.1016/j.neuron.2024.06.002. Epub 2024 Jul 29.

Authors

Isabel Lam¹, Alain Ndayisaba², Amanda J Lewis³, YuHong Fu⁴, Giselle T Sagredo⁴, Anastasia Kuzkina¹, Ludovica Zaccagnini⁵, Meral Celikag⁵, Jackson Sandoe⁶, Ricardo L Sanz¹, Aazam Vahdatshoar⁷, Timothy D Martin⁸, Nader Morshed⁹, Toru Ichihashi¹⁰, Arati Tripathi¹¹, Nagendran Ramalingam¹¹, Charlotte Oettgen-Suazo⁷, Theresa Bartels⁶, Manel Boussouf⁷, Max Schäbinger⁷, Erinc Hallacli¹, Xin Jiang¹², Amrita Verma⁷, Challana Tea¹³, Zichen Wang¹³, Hiroyuki Hakozaki¹³, Xiao Yu⁷, Kelly Hyles⁷, Chansaem Park⁷, Xinyuan Wang¹, Thorold W Theunissen⁶, Haoyi Wang⁶, Rudolf Jaenisch⁶, Susan Lindquist¹⁴, Beth Stevens⁹, Nadia Stefanova¹⁵, Gregor Wenning¹⁵, Wilma D J van de Berg¹⁶, Kelvin C Luk¹⁷, Rosario Sanchez-Pernaute¹⁸, Juan Carlos Gómez-Esteban¹⁹, Daniel Felsky²⁰, Yasujiro Kiyota¹⁰, Nidhi Sahni²¹, S Stephen Yi²², Chee Yeun Chung¹², Henning Stahlberg³, Isidro Ferrer²³, Johannes Schöneberg¹³, Stephen J Elledge⁸, Ulf Dettmer¹¹, Glenda M Halliday²⁴, Tim Bartels⁵, Vikram Khurana²⁵

Affiliations

¹ Ann Romney Center for Neurologic Diseases, Brigham and Women's Hospital, Boston, MA, USA; Division of Movement Disorders, American Parkinson Disease Association (APDA) Center for Advanced Research and MSA Center of Excellence, Department of Neurology, Brigham and Women's Hospital, Boston, MA, USA; Harvard Medical School, Boston, MA, USA.
² Ann Romney Center for Neurologic Diseases, Brigham and Women's Hospital, Boston, MA, USA; Division of Movement Disorders, American Parkinson Disease Association (APDA) Center for Advanced Research and MSA Center of Excellence, Department of Neurology, Brigham and Women's Hospital, Boston, MA, USA; Harvard Medical School, Boston, MA, USA; Division of Neurobiology, Department of Neurology, Medical University of Innsbruck, Innsbruck, Austria.
³ École Polytechnique Fédérale de Lausanne and University of Lausanne, Lausanne, Switzerland.
⁴ The University of Sydney Brain and Mind Centre and Faculty of Medicine and Health School of Medical Science, Sydney, NSW, Australia.
⁵ Dementia Research Institute, University College London, London, UK.
⁶ Whitehead Institute for Biomedical Research, Cambridge, MA, USA.
⁷ Ann Romney Center for Neurologic Diseases, Brigham and Women's Hospital, Boston, MA, USA; Division of Movement Disorders, American Parkinson Disease Association (APDA) Center for Advanced Research and MSA Center of Excellence, Department of Neurology, Brigham and Women's Hospital, Boston, MA, USA.
⁸ Harvard Medical School, Boston, MA, USA; Division of Genetics, Department of Medicine, Brigham and Women's Hospital, Boston, MA, USA; Howard Hughes Medical Institute, Chevy Chase, MD, USA.
⁹ Harvard Medical School, Boston, MA, USA; Howard Hughes Medical Institute, Chevy Chase, MD, USA; Boston Children's Hospital, Boston, MA, USA; The Broad Institute of MIT and Harvard, Cambridge, MA, USA.
¹⁰ Nikon Corporation, Tokyo, Japan.
¹¹ Ann Romney Center for Neurologic Diseases, Brigham and Women's Hospital, Boston, MA, USA; Harvard Medical School, Boston, MA, USA.
¹² Yumanity Therapeutics, Cambridge, MA, USA.
¹³ University of California, San Diego, San Diego, CA, USA.
¹⁴ Whitehead Institute for Biomedical Research, Cambridge, MA, USA; Howard Hughes Medical Institute, Chevy Chase, MD, USA.
¹⁵ Division of Neurobiology, Department of Neurology, Medical University of Innsbruck, Innsbruck, Austria.
¹⁶ Amsterdam University Medical Center, Vrije Universiteit, Amsterdam, the Netherlands.
¹⁷ University of Pennsylvania Perelman School of Medicine, Philadelphia, PA, USA.
¹⁸ BioBizkaia Health Research Institute, Barakaldo, Spain; Ikerbasque, Basque Foundation for Science, Bilbao, Spain.
¹⁹ BioBizkaia Health Research Institute, Barakaldo, Spain.
²⁰ Centre for Addiction and Mental Health, Toronto, ON, Canada; University of Toronto, Toronto, ON, Canada.
²¹ The University of Texas MD Anderson Cancer Center, Houston, TX, USA; Baylor College of Medicine, Houston, TX, USA.
²² The University of Texas at Austin, Austin, TX, USA.
²³ The University of Barcelona, Institut d'Investigacio Biomedica de Bellvitge IDIBELL, Hospitalet de Llobregat, Barcelona, Spain.
²⁴ The University of Sydney Brain and Mind Centre and Faculty of Medicine and Health School of Medical Science, Sydney, NSW, Australia; Aligning Science Across Parkinson's (ASAP) Collaborative Research Network, Chevy Chase, MD, USA.
²⁵ Ann Romney Center for Neurologic Diseases, Brigham and Women's Hospital, Boston, MA, USA; Division of Movement Disorders, American Parkinson Disease Association (APDA) Center for Advanced Research and MSA Center of Excellence, Department of Neurology, Brigham and Women's Hospital, Boston, MA, USA; Harvard Medical School, Boston, MA, USA; The Broad Institute of MIT and Harvard, Cambridge, MA, USA; Aligning Science Across Parkinson's (ASAP) Collaborative Research Network, Chevy Chase, MD, USA; Harvard Stem Cell Institute, Cambridge, MA, USA. Electronic address: khuranalab_admin@bwh.harvard.edu.

Abstract

The heterogeneity of protein-rich inclusions and its significance in neurodegeneration is poorly understood. Standard patient-derived iPSC models develop inclusions neither reproducibly nor in a reasonable time frame. Here, we developed screenable iPSC "inclusionopathy" models utilizing piggyBac or targeted transgenes to rapidly induce CNS cells that express aggregation-prone proteins at brain-like levels. Inclusions and their effects on cell survival were trackable at single-inclusion resolution. Exemplar cortical neuron α-synuclein inclusionopathy models were engineered through transgenic expression of α-synuclein mutant forms or exogenous seeding with fibrils. We identified multiple inclusion classes, including neuroprotective p62-positive inclusions versus dynamic and neurotoxic lipid-rich inclusions, both identified in patient brains. Fusion events between these inclusion subtypes altered neuronal survival. Proteome-scale α-synuclein genetic- and physical-interaction screens pinpointed candidate RNA-processing and actin-cytoskeleton-modulator proteins like RhoA whose sequestration into inclusions could enhance toxicity. These tractable CNS models should prove useful in functional genomic analysis and drug development for proteinopathies.

Keywords: CRISPR screen; Lewy body; Parkinson’s disease; Rab protein; RhoA; actin cytoskeleton; aggregation; dementia with Lewy bodies; glia; iPSC; inclusion; lipid; neurodegeneration; neuron; p62; piggyBac; proximity labeling; synucleinopathy; ubiquitin; α-synuclein.

MeSH terms

Brain / metabolism
Brain / pathology
Humans
Inclusion Bodies* / metabolism
Inclusion Bodies* / pathology
Induced Pluripotent Stem Cells* / metabolism
Neurons / metabolism
Neurons / pathology
Synucleinopathies / genetics
Synucleinopathies / metabolism
Synucleinopathies / pathology
alpha-Synuclein* / genetics
alpha-Synuclein* / metabolism

Substances

alpha-Synuclein

Abstract

MeSH terms

Substances

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