CryoET reveals organelle phenotypes in huntington disease patient iPSC-derived and mouse primary neurons

Gong-Her Wu; Charlene Smith-Geater; Jesús G Galaz-Montoya; Yingli Gu; Sanket R Gupte; Ranen Aviner; Patrick G Mitchell; Joy Hsu; Ricardo Miramontes; Keona Q Wang; Nicolette R Geller; Cathy Hou; Cristina Danita; Lydia-Marie Joubert; Michael F Schmid; Serena Yeung; Judith Frydman; William Mobley; Chengbiao Wu; Leslie M Thompson; Wah Chiu

doi:10.1038/s41467-023-36096-w

CryoET reveals organelle phenotypes in huntington disease patient iPSC-derived and mouse primary neurons

Nat Commun. 2023 Feb 8;14(1):692. doi: 10.1038/s41467-023-36096-w.

Authors

Gong-Her Wu^#¹, Charlene Smith-Geater^#², Jesús G Galaz-Montoya¹, Yingli Gu³, Sanket R Gupte⁴, Ranen Aviner⁵, Patrick G Mitchell⁶, Joy Hsu⁴, Ricardo Miramontes⁷, Keona Q Wang⁸, Nicolette R Geller⁸, Cathy Hou¹, Cristina Danita¹, Lydia-Marie Joubert⁶, Michael F Schmid⁶, Serena Yeung^{4

9}, Judith Frydman^{5

10}, William Mobley³, Chengbiao Wu³, Leslie M Thompson^{11

12

13

14

15}, Wah Chiu^{16

17

18}

Affiliations

¹ Department of Bioengineering, James H. Clark Center, Stanford University, Stanford, CA, 94305, USA.
² Department of Psychiatry & Human Behavior University of California Irvine, Irvine, CA, 92697, USA.
³ Department of Neurosciences, University of California San Diego, La Jolla, CA, 92037-0662, USA.
⁴ Department of Computer Science, Stanford University, Stanford, CA, 94305, USA.
⁵ Department of Biology, Stanford University, Stanford, CA, 94305, USA.
⁶ Division of CryoEM and Bioimaging, SSRL, SLAC National Accelerator Laboratory, Stanford University, Menlo Park, CA, 94025, USA.
⁷ Department of Memory Impairment and Neurological Disorders, University of California Irvine, Irvine, CA, 92697, USA.
⁸ Department of Neurobiology and Behavior, University of California Irvine, Irvine, CA, 96267, USA.
⁹ Department of Biomedical Data Science, Stanford University, Stanford, CA, 94305, USA.
¹⁰ Department of Genetics, Stanford University, Stanford, CA, 94305, USA.
¹¹ Department of Psychiatry & Human Behavior University of California Irvine, Irvine, CA, 92697, USA. lmthomps@uci.edu.
¹² Department of Memory Impairment and Neurological Disorders, University of California Irvine, Irvine, CA, 92697, USA. lmthomps@uci.edu.
¹³ Department of Neurobiology and Behavior, University of California Irvine, Irvine, CA, 96267, USA. lmthomps@uci.edu.
¹⁴ Sue & Bill Gross Stem Cell Research Center, University of California Irvine, Irvine, CA, 96267, USA. lmthomps@uci.edu.
¹⁵ Department of Biological Chemistry, University of California Irvine, Irvine, CA, 92617, USA. lmthomps@uci.edu.
¹⁶ Department of Bioengineering, James H. Clark Center, Stanford University, Stanford, CA, 94305, USA. wahc@stanford.edu.
¹⁷ Division of CryoEM and Bioimaging, SSRL, SLAC National Accelerator Laboratory, Stanford University, Menlo Park, CA, 94025, USA. wahc@stanford.edu.
¹⁸ Department of Microbiology and Immunology, Stanford University, Stanford, CA, 94305, USA. wahc@stanford.edu.

^# Contributed equally.

Abstract

Huntington's disease (HD) is caused by an expanded CAG repeat in the huntingtin gene, yielding a Huntingtin protein with an expanded polyglutamine tract. While experiments with patient-derived induced pluripotent stem cells (iPSCs) can help understand disease, defining pathological biomarkers remains challenging. Here, we used cryogenic electron tomography to visualize neurites in HD patient iPSC-derived neurons with varying CAG repeats, and primary cortical neurons from BACHD, deltaN17-BACHD, and wild-type mice. In HD models, we discovered sheet aggregates in double membrane-bound organelles, and mitochondria with distorted cristae and enlarged granules, likely mitochondrial RNA granules. We used artificial intelligence to quantify mitochondrial granules, and proteomics experiments reveal differential protein content in isolated HD mitochondria. Knockdown of Protein Inhibitor of Activated STAT1 ameliorated aberrant phenotypes in iPSC- and BACHD neurons. We show that integrated ultrastructural and proteomic approaches may uncover early HD phenotypes to accelerate diagnostics and the development of targeted therapeutics for HD.

Publication types

Research Support, Non-U.S. Gov't
Research Support, N.I.H., Extramural

MeSH terms

Animals
Artificial Intelligence
Disease Models, Animal
Humans
Huntingtin Protein / genetics
Huntingtin Protein / metabolism
Huntington Disease* / metabolism
Induced Pluripotent Stem Cells* / metabolism
Mice
Mitochondria / metabolism
Neurons / metabolism
Phenotype
Proteomics

Substances

Huntingtin Protein

Abstract

Publication types

MeSH terms

Substances

Grants and funding