Chimeric Peptide Species Contribute to Divergent Dipeptide Repeat Pathology in c9ALS/FTD and SCA36

Neuro–CEB Neuropathology Network

doi:10.1016/j.neuron.2020.04.011

Chimeric Peptide Species Contribute to Divergent Dipeptide Repeat Pathology in c9ALS/FTD and SCA36

Neuro–CEB Neuropathology Network

Research output: Contribution to journal › Article › peer-review

37 Citations (Scopus)

Abstract

GGGGCC hexanucleotide repeat expansions (HREs) in C9orf72 cause amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD) and lead to the production of aggregating dipeptide repeat proteins (DPRs) via repeat associated non-AUG (RAN) translation. Here, we show the similar intronic GGCCTG HREs that causes spinocerebellar ataxia type 36 (SCA36) is also translated into DPRs, including poly(GP) and poly(PR). We demonstrate that poly(GP) is more abundant in SCA36 compared to c9ALS/FTD patient tissue due to canonical AUG-mediated translation from intron-retained GGCCTG repeat RNAs. However, the frequency of the antisense RAN translation product poly(PR) is comparable between c9ALS/FTD and SCA36 patient samples. Interestingly, in SCA36 patient tissue, poly(GP) exists as a soluble species, and no TDP-43 pathology is present. We show that aggregate-prone chimeric DPR (cDPR) species underlie the divergent DPR pathology between c9ALS/FTD and SCA36. These findings reveal key differences in translation, solubility, and protein aggregation of DPRs between c9ALS/FTD and SCA36.

Original language	English
Pages (from-to)	292-305.e6
Journal	Neuron
Volume	107
Issue number	2
DOIs	https://doi.org/10.1016/j.neuron.2020.04.011
Publication status	Published - Jul 22 2020

Keywords

ALS
C9orf72 expansion
FTD
RAN translation
SCA36
antisense oligonucleotide therapy
chimeric DPRs
dipeptide repeats
neurodegeneration

ASJC Scopus subject areas

Neuroscience(all)

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10.1016/j.neuron.2020.04.011

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@article{909810de4f28400aa333edd59be92ab7,

title = "Chimeric Peptide Species Contribute to Divergent Dipeptide Repeat Pathology in c9ALS/FTD and SCA36",

abstract = "GGGGCC hexanucleotide repeat expansions (HREs) in C9orf72 cause amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD) and lead to the production of aggregating dipeptide repeat proteins (DPRs) via repeat associated non-AUG (RAN) translation. Here, we show the similar intronic GGCCTG HREs that causes spinocerebellar ataxia type 36 (SCA36) is also translated into DPRs, including poly(GP) and poly(PR). We demonstrate that poly(GP) is more abundant in SCA36 compared to c9ALS/FTD patient tissue due to canonical AUG-mediated translation from intron-retained GGCCTG repeat RNAs. However, the frequency of the antisense RAN translation product poly(PR) is comparable between c9ALS/FTD and SCA36 patient samples. Interestingly, in SCA36 patient tissue, poly(GP) exists as a soluble species, and no TDP-43 pathology is present. We show that aggregate-prone chimeric DPR (cDPR) species underlie the divergent DPR pathology between c9ALS/FTD and SCA36. These findings reveal key differences in translation, solubility, and protein aggregation of DPRs between c9ALS/FTD and SCA36.",

keywords = "ALS, C9orf72 expansion, FTD, RAN translation, SCA36, antisense oligonucleotide therapy, chimeric DPRs, dipeptide repeats, neurodegeneration",

author = "{Neuro–CEB Neuropathology Network} and McEachin, {Zachary T.} and Gendron, {Tania F.} and Nisha Raj and Mar{\'i}a Garc{\'i}a-Murias and Anwesha Banerjee and Purcell, {Ryan H.} and Ward, {Patricia J.} and Todd, {Tiffany W.} and Merritt-Garza, {Megan E.} and Karen Jansen-West and Hales, {Chadwick M.} and Tania Garc{\'i}a-Sobrino and Beatriz Quint{\'a}ns and Holler, {Christopher J.} and Georgia Taylor and {San Mill{\'a}n}, Beatriz and Susana Teijeira and Toru Yamashita and Ryuichi Ohkubo and Boulis, {Nicholas M.} and Chongchong Xu and Zhexing Wen and Nathalie Streichenberger and Fogel, {Brent L.} and Thomas Kukar and Koji Abe and Dickson, {Dennis W.} and Manuel Arias and Glass, {Jonathan D.} and Jie Jiang and Tansey, {Mal{\'u} G.} and Sobrido, {Mar{\'i}a Jes{\'u}s} and Leonard Petrucelli and Wilfried Rossoll and Bassell, {Gary J.}",

note = "Funding Information: We are extremely grateful to the patients and their families for participating in this study. We thank the Galician Ataxia Assocation (AGA) for their cooperation and continuing support. We would also like to acknowledge the technical expertise provided by Marla Gearing, Deborah Cooper, and Kaylor Kelly of the Emory Neuropathology Core. We would like to acknowledge Alexandra D?rr of Piti?-Salp?tri?re Hospital - Sorbonne Universit? for her assistance in obtaining patient samples. Additionally, we would like to thank Dr. Dieter Edbauer for kindly providing the monoclonal clone 5F2 poly(GA) antibody (provided to J.J.). We would like to acknowledge Yan Liang, Jingjing Gong, and Liang Zhang for their assistance with DSP data collection and analysis. Human biological samples and associated data were obtained from the Emory Neuropathology Core (P30 NS055077), Brain Bank of Biobank Galicia Sur Health Research Institute (PT17/0015/0034), Brainbank Neuro-CEB, H?pital de la Piti?-Salp?tri?re, and Tissu-Tumorotheque Est, Hospices Civils de Lyon Biobank (CRB-HCL, BB-0033-00046). This work was supported by an Emory University Research Committee grant (G.J.B.), Emory University School of Medicine Laboratory for Translational Cell Biology (G.J.B), National Institute for Neurological Disorders and Stroke grant R21NS114908 (J.J. and G.J.B), the Center for Neurodysfunction and Inflammation at Emory (M.G.T. and G.J.B.), the National Ataxia Foundation (W.R.), National Institute for Neurological Disorders and Stroke grant R01NS91749 (W.R.), National Institute for Neurological Disorders and Stroke grant R01NS082094 (B.L.F.), the National Institutes of Health/National Institute for Neurological Disorders and Stroke grants R35NS097273 (L.P.), P01NS084974 (L.P. and D.W.D.), and P01NS099114 (T.F.G. and L.P.), and ISCIII grants PI12/00742 and PI17/01789 (M.-J.S.), cofounded by FEDER. Z.T.M. was supported by a Training and Research in Translational Neurology training grant (2T32 NS007480-15). Z.T.M. conceived and coordinated the project. Z.T.M. T.F.G. L.P. W.R. and G.J.B. designed experiments. Z.T.M. T.F.G. J.J. W.R. and G.J.B. analyzed and interpreted data. Z.T.M. generated iPSCs; performed iPSC differentiations, immunohistochemistry in patient tissue, and ASO experiments; and prepared protein lysates, western blot, qPCR, immunofluorescence staining, luciferase assays, RP-PCR, and sequencing. T.F.G. performed MSD immunoassays. T.F.G. and D.W.D. performed poly(PR) immunohistochemistry. Z.T.M. and K.J-W. made constructs. Z.T.M. N.R. M.E.M.-G. and M.G.-M. performed cell culture. C.X. and Z.W. performed organoid differentiations. A.B. performed ICV injections. Z.T.M. A.B. and P.J.W. processed mice tissue and performed immunostaining. Z.T.M. and J.J. performed poly(GP)/poly(GA) co-localization experiment. Z.T.M. J.J. and M.G.T. analyzed DSP data. R.H.P. N.B. C.H. G.T. T.W.T. T.K. M.G.T. and J.D.G. provided technical support. C.M.H. B.S.M. T.G.-S. B.Q. S.T. T.Y. R.O. B.L.F. N.S. K.A. M.A. J.D.G. and M.-J.S. provided patient samples and associated genetic and clinical data. Z.T.M. wrote the manuscript. Z.T.M. T.F.G. N.R. J.J. M.G.T. M.A. J.D.G. M.-J.S. W.R. and G.J.B. revised and edited the manuscript. The authors declare no competing interests. Funding Information: We are extremely grateful to the patients and their families for participating in this study. We thank the Galician Ataxia Assocation (AGA) for their cooperation and continuing support. We would also like to acknowledge the technical expertise provided by Marla Gearing, Deborah Cooper, and Kaylor Kelly of the Emory Neuropathology Core. We would like to acknowledge Alexandra D{\"u}rr of Piti{\'e}-Salp{\^e}tri{\`e}re Hospital - Sorbonne Universit{\'e} for her assistance in obtaining patient samples. Additionally, we would like to thank Dr. Dieter Edbauer for kindly providing the monoclonal clone 5F2 poly(GA) antibody (provided to J.J.). We would like to acknowledge Yan Liang, Jingjing Gong, and Liang Zhang for their assistance with DSP data collection and analysis. Human biological samples and associated data were obtained from the Emory Neuropathology Core ( P30 NS055077 ), Brain Bank of Biobank Galicia Sur Health Research Institute ( PT17/0015/0034 ), Brainbank Neuro-CEB, H{\^o}pital de la Piti{\'e}-Salp{\^e}tri{\`e}re, and Tissu-Tumorotheque Est, Hospices Civils de Lyon Biobank (CRB-HCL, BB-0033-00046 ). This work was supported by an Emory University Research Committee grant (G.J.B.), Emory University School of Medicine Laboratory for Translational Cell Biology (G.J.B), National Institute for Neurological Disorders and Stroke grant R21NS114908 (J.J. and G.J.B), the Center for Neurodysfunction and Inflammation at Emory (M.G.T. and G.J.B.), the National Ataxia Foundation (W.R.), National Institute for Neurological Disorders and Stroke grant R01NS91749 (W.R.), National Institute for Neurological Disorders and Stroke grant R01NS082094 (B.L.F.), the National Institutes of Health / National Institute for Neurological Disorders and Stroke grants R35NS097273 (L.P.), P01NS084974 (L.P. and D.W.D.), and P01NS099114 (T.F.G. and L.P.), and ISCIII grants PI12/00742 and PI17/01789 (M.-J.S.), cofounded by FEDER . Z.T.M. was supported by a Training and Research in Translational Neurology training grant ( 2T32 NS007480-15 ). Publisher Copyright: {\textcopyright} 2020 Elsevier Inc.",

year = "2020",

month = jul,

day = "22",

doi = "10.1016/j.neuron.2020.04.011",

language = "English",

volume = "107",

pages = "292--305.e6",

journal = "Neuron",

issn = "0896-6273",

publisher = "Cell Press",

number = "2",

}

TY - JOUR

T1 - Chimeric Peptide Species Contribute to Divergent Dipeptide Repeat Pathology in c9ALS/FTD and SCA36

AU - Neuro–CEB Neuropathology Network

AU - McEachin, Zachary T.

AU - Gendron, Tania F.

AU - Raj, Nisha

AU - García-Murias, María

AU - Banerjee, Anwesha

AU - Purcell, Ryan H.

AU - Ward, Patricia J.

AU - Todd, Tiffany W.

AU - Merritt-Garza, Megan E.

AU - Jansen-West, Karen

AU - Hales, Chadwick M.

AU - García-Sobrino, Tania

AU - Quintáns, Beatriz

AU - Holler, Christopher J.

AU - Taylor, Georgia

AU - San Millán, Beatriz

AU - Teijeira, Susana

AU - Yamashita, Toru

AU - Ohkubo, Ryuichi

AU - Boulis, Nicholas M.

AU - Xu, Chongchong

AU - Wen, Zhexing

AU - Streichenberger, Nathalie

AU - Fogel, Brent L.

AU - Kukar, Thomas

AU - Abe, Koji

AU - Dickson, Dennis W.

AU - Arias, Manuel

AU - Glass, Jonathan D.

AU - Jiang, Jie

AU - Tansey, Malú G.

AU - Sobrido, María Jesús

AU - Petrucelli, Leonard

AU - Rossoll, Wilfried

AU - Bassell, Gary J.

N1 - Funding Information: We are extremely grateful to the patients and their families for participating in this study. We thank the Galician Ataxia Assocation (AGA) for their cooperation and continuing support. We would also like to acknowledge the technical expertise provided by Marla Gearing, Deborah Cooper, and Kaylor Kelly of the Emory Neuropathology Core. We would like to acknowledge Alexandra D?rr of Piti?-Salp?tri?re Hospital - Sorbonne Universit? for her assistance in obtaining patient samples. Additionally, we would like to thank Dr. Dieter Edbauer for kindly providing the monoclonal clone 5F2 poly(GA) antibody (provided to J.J.). We would like to acknowledge Yan Liang, Jingjing Gong, and Liang Zhang for their assistance with DSP data collection and analysis. Human biological samples and associated data were obtained from the Emory Neuropathology Core (P30 NS055077), Brain Bank of Biobank Galicia Sur Health Research Institute (PT17/0015/0034), Brainbank Neuro-CEB, H?pital de la Piti?-Salp?tri?re, and Tissu-Tumorotheque Est, Hospices Civils de Lyon Biobank (CRB-HCL, BB-0033-00046). This work was supported by an Emory University Research Committee grant (G.J.B.), Emory University School of Medicine Laboratory for Translational Cell Biology (G.J.B), National Institute for Neurological Disorders and Stroke grant R21NS114908 (J.J. and G.J.B), the Center for Neurodysfunction and Inflammation at Emory (M.G.T. and G.J.B.), the National Ataxia Foundation (W.R.), National Institute for Neurological Disorders and Stroke grant R01NS91749 (W.R.), National Institute for Neurological Disorders and Stroke grant R01NS082094 (B.L.F.), the National Institutes of Health/National Institute for Neurological Disorders and Stroke grants R35NS097273 (L.P.), P01NS084974 (L.P. and D.W.D.), and P01NS099114 (T.F.G. and L.P.), and ISCIII grants PI12/00742 and PI17/01789 (M.-J.S.), cofounded by FEDER. Z.T.M. was supported by a Training and Research in Translational Neurology training grant (2T32 NS007480-15). Z.T.M. conceived and coordinated the project. Z.T.M. T.F.G. L.P. W.R. and G.J.B. designed experiments. Z.T.M. T.F.G. J.J. W.R. and G.J.B. analyzed and interpreted data. Z.T.M. generated iPSCs; performed iPSC differentiations, immunohistochemistry in patient tissue, and ASO experiments; and prepared protein lysates, western blot, qPCR, immunofluorescence staining, luciferase assays, RP-PCR, and sequencing. T.F.G. performed MSD immunoassays. T.F.G. and D.W.D. performed poly(PR) immunohistochemistry. Z.T.M. and K.J-W. made constructs. Z.T.M. N.R. M.E.M.-G. and M.G.-M. performed cell culture. C.X. and Z.W. performed organoid differentiations. A.B. performed ICV injections. Z.T.M. A.B. and P.J.W. processed mice tissue and performed immunostaining. Z.T.M. and J.J. performed poly(GP)/poly(GA) co-localization experiment. Z.T.M. J.J. and M.G.T. analyzed DSP data. R.H.P. N.B. C.H. G.T. T.W.T. T.K. M.G.T. and J.D.G. provided technical support. C.M.H. B.S.M. T.G.-S. B.Q. S.T. T.Y. R.O. B.L.F. N.S. K.A. M.A. J.D.G. and M.-J.S. provided patient samples and associated genetic and clinical data. Z.T.M. wrote the manuscript. Z.T.M. T.F.G. N.R. J.J. M.G.T. M.A. J.D.G. M.-J.S. W.R. and G.J.B. revised and edited the manuscript. The authors declare no competing interests. Funding Information: We are extremely grateful to the patients and their families for participating in this study. We thank the Galician Ataxia Assocation (AGA) for their cooperation and continuing support. We would also like to acknowledge the technical expertise provided by Marla Gearing, Deborah Cooper, and Kaylor Kelly of the Emory Neuropathology Core. We would like to acknowledge Alexandra Dürr of Pitié-Salpêtrière Hospital - Sorbonne Université for her assistance in obtaining patient samples. Additionally, we would like to thank Dr. Dieter Edbauer for kindly providing the monoclonal clone 5F2 poly(GA) antibody (provided to J.J.). We would like to acknowledge Yan Liang, Jingjing Gong, and Liang Zhang for their assistance with DSP data collection and analysis. Human biological samples and associated data were obtained from the Emory Neuropathology Core ( P30 NS055077 ), Brain Bank of Biobank Galicia Sur Health Research Institute ( PT17/0015/0034 ), Brainbank Neuro-CEB, Hôpital de la Pitié-Salpêtrière, and Tissu-Tumorotheque Est, Hospices Civils de Lyon Biobank (CRB-HCL, BB-0033-00046 ). This work was supported by an Emory University Research Committee grant (G.J.B.), Emory University School of Medicine Laboratory for Translational Cell Biology (G.J.B), National Institute for Neurological Disorders and Stroke grant R21NS114908 (J.J. and G.J.B), the Center for Neurodysfunction and Inflammation at Emory (M.G.T. and G.J.B.), the National Ataxia Foundation (W.R.), National Institute for Neurological Disorders and Stroke grant R01NS91749 (W.R.), National Institute for Neurological Disorders and Stroke grant R01NS082094 (B.L.F.), the National Institutes of Health / National Institute for Neurological Disorders and Stroke grants R35NS097273 (L.P.), P01NS084974 (L.P. and D.W.D.), and P01NS099114 (T.F.G. and L.P.), and ISCIII grants PI12/00742 and PI17/01789 (M.-J.S.), cofounded by FEDER . Z.T.M. was supported by a Training and Research in Translational Neurology training grant ( 2T32 NS007480-15 ). Publisher Copyright: © 2020 Elsevier Inc.

PY - 2020/7/22

Y1 - 2020/7/22

N2 - GGGGCC hexanucleotide repeat expansions (HREs) in C9orf72 cause amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD) and lead to the production of aggregating dipeptide repeat proteins (DPRs) via repeat associated non-AUG (RAN) translation. Here, we show the similar intronic GGCCTG HREs that causes spinocerebellar ataxia type 36 (SCA36) is also translated into DPRs, including poly(GP) and poly(PR). We demonstrate that poly(GP) is more abundant in SCA36 compared to c9ALS/FTD patient tissue due to canonical AUG-mediated translation from intron-retained GGCCTG repeat RNAs. However, the frequency of the antisense RAN translation product poly(PR) is comparable between c9ALS/FTD and SCA36 patient samples. Interestingly, in SCA36 patient tissue, poly(GP) exists as a soluble species, and no TDP-43 pathology is present. We show that aggregate-prone chimeric DPR (cDPR) species underlie the divergent DPR pathology between c9ALS/FTD and SCA36. These findings reveal key differences in translation, solubility, and protein aggregation of DPRs between c9ALS/FTD and SCA36.

AB - GGGGCC hexanucleotide repeat expansions (HREs) in C9orf72 cause amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD) and lead to the production of aggregating dipeptide repeat proteins (DPRs) via repeat associated non-AUG (RAN) translation. Here, we show the similar intronic GGCCTG HREs that causes spinocerebellar ataxia type 36 (SCA36) is also translated into DPRs, including poly(GP) and poly(PR). We demonstrate that poly(GP) is more abundant in SCA36 compared to c9ALS/FTD patient tissue due to canonical AUG-mediated translation from intron-retained GGCCTG repeat RNAs. However, the frequency of the antisense RAN translation product poly(PR) is comparable between c9ALS/FTD and SCA36 patient samples. Interestingly, in SCA36 patient tissue, poly(GP) exists as a soluble species, and no TDP-43 pathology is present. We show that aggregate-prone chimeric DPR (cDPR) species underlie the divergent DPR pathology between c9ALS/FTD and SCA36. These findings reveal key differences in translation, solubility, and protein aggregation of DPRs between c9ALS/FTD and SCA36.

KW - ALS

KW - C9orf72 expansion

KW - FTD

KW - RAN translation

KW - SCA36

KW - antisense oligonucleotide therapy

KW - chimeric DPRs

KW - dipeptide repeats

KW - neurodegeneration

UR - http://www.scopus.com/inward/record.url?scp=85084974096&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85084974096&partnerID=8YFLogxK

U2 - 10.1016/j.neuron.2020.04.011

DO - 10.1016/j.neuron.2020.04.011

M3 - Article

C2 - 32375063

AN - SCOPUS:85084974096

SN - 0896-6273

VL - 107

SP - 292-305.e6

JO - Neuron

JF - Neuron

IS - 2

ER -

Chimeric Peptide Species Contribute to Divergent Dipeptide Repeat Pathology in c9ALS/FTD and SCA36

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