<i>CASP8</i> intronic expansion identified by poly-glycine-arginine pathology increases Alzheimer’s disease risk

L Lien Nguyen R Ramadan Ajredini (Center for NeuroGenetics, College of Medicine, University of Florida) S Shu Guo (Shenzhen Institute for Quantum Science and Engineering, Department of Chemistry, and Department of Physics) L Lisa E. L. Romano (Center for NeuroGenetics, College of Medicine, University of Florida) R Rodrigo F. Tomas (Center for NeuroGenetics, College of Medicine, University of Florida) L Logan R. Bell (Center for NeuroGenetics, College of Medicine, University of Florida) P Paul T. Ranum (Raymond G. Perelman Center for Cellular and Molecular Therapeutics, The Children’s Hospital of Philadelphia, University of Pennsylvania) T Tao Zu (Center for NeuroGenetics, College of Medicine, University of Florida) M Monica Bañez Coronel (Center for NeuroGenetics, College of Medicine, University of Florida) C Chase P. Kelley (Center for NeuroGenetics, College of Medicine, University of Florida) J Javier Redding-Ochoa (Department of Pathology, The Johns Hopkins University School of Medicine) E Evangelos Nizamis (Division of Medical Genetics School of Medicine, University of Washington) A Alexandra Melloni (Department of Neurology, MassGeneral Institute for Neurodegenerative Disease, Massachusetts General Hospital) T Theresa R. Connors (Department of Neurology, MassGeneral Institute for Neurodegenerative Disease, Massachusetts General Hospital) A Angelica Gaona (Department of Neurology, MassGeneral Institute for Neurodegenerative Disease, Massachusetts General Hospital) K Kiruphagaran Thangaraju (Center for NeuroGenetics, College of Medicine, University of Florida) O Olga Pletnikova (Department of Pathology, The Johns Hopkins University School of Medicine) H H. Brent Clark (Department of Laboratory Medicine and Pathology, University of Minnesota) B Beverly L. Davidson (Raymond G. Perelman Center for Cellular and Molecular Therapeutics, The Children’s Hospital of Philadelphia, University of Pennsylvania) A Anthony T. Yachnis (Center for NeuroGenetics, College of Medicine, University of Florida) T Todd E. Golde (Center for Translation Research in Neurodegenerative Disease, College of Medicine, University of Florida) X XiangYang Lou (Department of Biostatistics, University of Florida) E Eric T. Wang (Center for NeuroGenetics, College of Medicine, University of Florida) A Alan E. Renton (Ronald M. Loeb Center for Alzheimer’s Disease, Department of Genetics and Genomic Sciences, Icahn School of Medicine at Mount Sinai) A Alison Goate (Ronald M. Loeb Center for Alzheimer’s Disease, Department of Genetics and Genomic Sciences, Icahn School of Medicine at Mount Sinai) P Paul N. Valdmanis (Division of Medical Genetics School of Medicine, University of Washington) S Stefan Prokop (Center for NeuroGenetics, College of Medicine, University of Florida) J Juan C. Troncoso (Department of Pathology, The Johns Hopkins University School of Medicine) B Bradley T. Hyman L Laura P. W. Ranum (Center for NeuroGenetics, College of Medicine, University of Florida)

Abstract

Alzheimer’s disease (AD) affects more than 10% of the population ≥65 y of age, but the underlying biological risks of most AD cases are unclear. We show anti-poly-glycine-arginine (a-polyGR) positive aggregates frequently accumulate in sporadic AD autopsy brains (45/80 cases). We hypothesize that these aggregates are caused by one or more polyGR-encoding repeat expansion mutations. We developed a CRISPR/deactivated-Cas9 enrichment strategy to identify candidate GR-encoding repeat expansion mutations directly from genomic DNA isolated from a-polyGR(+) AD cases. Using this approach, we isolated an interrupted (GGGAGA) n intronic expansion within a SINE-VNTR-Alu element in CASP8 ( CASP8 -GGGAGA EXP ). Immunostaining using a-polyGR and locus-specific C-terminal antibodies demonstrate that the CASP8 -GGGAGA EXP expresses hybrid poly(GR)n(GE)n(RE)n proteins that accumulate in CASP8 -GGGAGA EXP (+) AD brains. In cells, expression of CASP8 -GGGAGA EXP minigenes leads to increased p-Tau (Ser202/Thr205) levels. Consistent with other types of repeat-associated non-AUG (RAN) proteins, poly(GR)n(GE)n(RE)n protein levels are increased by stress. Additionally, levels of these stress-induced proteins are reduced by metformin. Association studies show specific aggregate promoting interrupted CASP8 -GGGAGA EXP sequence variants found in ~3.6% of controls and 7.5% AD cases increase AD risk [ CASP8 -GGGAGA-AD-R1; OR 2.2, 95% CI (1.5185 to 3.1896), P = 3.1 × 10 −5 ]. Cells transfected with a high-risk CASP8 -GGGAGA-AD-R1 variant show increased toxicity and increased levels of poly(GR)n(GE)n(RE)n aggregates. Taken together, these data identify polyGR(+) aggregates as a frequent and unexpected type of brain pathology in AD and CASP8 -GGGAGA-AD-R1 alleles as a relatively common AD risk factor. Taken together, these data support a model in which CASP8 -GGGAGA EXP alleles combined with stress increase AD risk.

Article Details

Volume / Issue Vol. 122, Issue 7
Published February 18, 2025
ISSN 0027-8424
Publisher National Academy of Sciences

Authors (30)

L

Lien Nguyen

R

Ramadan Ajredini

Center for NeuroGenetics, College of Medicine, University of Florida

S

Shu Guo

Shenzhen Institute for Quantum Science and Engineering, Department of Chemistry, and Department of Physics

L

Lisa E. L. Romano

Center for NeuroGenetics, College of Medicine, University of Florida

R

Rodrigo F. Tomas

Center for NeuroGenetics, College of Medicine, University of Florida

L

Logan R. Bell

Center for NeuroGenetics, College of Medicine, University of Florida

P

Paul T. Ranum

Raymond G. Perelman Center for Cellular and Molecular Therapeutics, The Children’s Hospital of Philadelphia, University of Pennsylvania

T

Tao Zu

Center for NeuroGenetics, College of Medicine, University of Florida

M

Monica Bañez Coronel

Center for NeuroGenetics, College of Medicine, University of Florida

C

Chase P. Kelley

Center for NeuroGenetics, College of Medicine, University of Florida

J

Javier Redding-Ochoa

Department of Pathology, The Johns Hopkins University School of Medicine

E

Evangelos Nizamis

Division of Medical Genetics School of Medicine, University of Washington

A

Alexandra Melloni

Department of Neurology, MassGeneral Institute for Neurodegenerative Disease, Massachusetts General Hospital

T

Theresa R. Connors

Department of Neurology, MassGeneral Institute for Neurodegenerative Disease, Massachusetts General Hospital

A

Angelica Gaona

Department of Neurology, MassGeneral Institute for Neurodegenerative Disease, Massachusetts General Hospital

K

Kiruphagaran Thangaraju

Center for NeuroGenetics, College of Medicine, University of Florida

O

Olga Pletnikova

Department of Pathology, The Johns Hopkins University School of Medicine

H

H. Brent Clark

Department of Laboratory Medicine and Pathology, University of Minnesota

B

Beverly L. Davidson

Raymond G. Perelman Center for Cellular and Molecular Therapeutics, The Children’s Hospital of Philadelphia, University of Pennsylvania

A

Anthony T. Yachnis

Center for NeuroGenetics, College of Medicine, University of Florida

T

Todd E. Golde

Center for Translation Research in Neurodegenerative Disease, College of Medicine, University of Florida

X

XiangYang Lou

Department of Biostatistics, University of Florida

E

Eric T. Wang

Center for NeuroGenetics, College of Medicine, University of Florida

A

Alan E. Renton

Ronald M. Loeb Center for Alzheimer’s Disease, Department of Genetics and Genomic Sciences, Icahn School of Medicine at Mount Sinai

A

Alison Goate

Ronald M. Loeb Center for Alzheimer’s Disease, Department of Genetics and Genomic Sciences, Icahn School of Medicine at Mount Sinai

P

Paul N. Valdmanis

Division of Medical Genetics School of Medicine, University of Washington

S

Stefan Prokop

Center for NeuroGenetics, College of Medicine, University of Florida

J

Juan C. Troncoso

Department of Pathology, The Johns Hopkins University School of Medicine

B

Bradley T. Hyman

L

Laura P. W. Ranum

Center for NeuroGenetics, College of Medicine, University of Florida