Characterization of HNRNPA1 mutations defines diversity in pathogenic mechanisms and clinical presentation.

Danique Beijer,Peter De Jonghe,James Shorter,Matthew T Wheeler,J Paul Taylor,Hong Joo Kim,Nicolas Dubuisson,Lauren E Drake,Peter Van Den Bergh,Tine Deconinck,Inès Mademan,Kristof Van Schil,Richard J.L.F Lemmers,Jonathan Baets,Silvère M Van Der Maarel,Devon Bonner,Jacinda B Sampson,Lin Guo,Nicol C Voermans,Dagmara Kabzińska,Anahit Mehrabyan,Andrzej Kochański,Charlotte M Fare,Alice Flynn Ford ,Kevin J O’donovan ,Steven C Palmer

doi:10.1172/jci.insight.148363

Abstract

Mutations in HNRNPA1 encoding heterogeneous nuclear ribonucleoprotein (hnRNP) A1 are a rare cause of amyotrophic lateral sclerosis (ALS) and multisystem proteinopathy (MSP). hnRNPA1 is part of the group of RNA-binding proteins (RBPs) that assemble with RNA to form RNPs. hnRNPs are concentrated in the nucleus and function in pre-mRNA splicing, mRNA stability, and the regulation of transcription and translation. During stress, hnRNPs, mRNA, and other RBPs condense in the cytoplasm to form stress granules (SGs). SGs are implicated in the pathogenesis of (neuro-)degenerative diseases, including ALS and inclusion body myopathy (IBM). Mutations in RBPs that affect SG biology, including FUS, TDP-43, hnRNPA1, hnRNPA2B1, and TIA1, underlie ALS, IBM, and other neurodegenerative diseases. Here, we characterize 4 potentially novel HNRNPA1 mutations (yielding 3 protein variants: *321Eext*6, *321Qext*6, and G304Nfs*3) and 2 known HNRNPA1 mutations (P288A and D262V), previously connected to ALS and MSP, in a broad spectrum of patients with hereditary motor neuropathy, ALS, and myopathy. We establish that the mutations can have different effects on hnRNPA1 fibrillization, liquid-liquid phase separation, and SG dynamics. P288A accelerated fibrillization and decelerated SG disassembly, whereas *321Eext*6 had no effect on fibrillization but decelerated SG disassembly. By contrast, G304Nfs*3 decelerated fibrillization and impaired liquid phase separation. Our findings suggest different underlying pathomechanisms for HNRNPA1 mutations with a possible link to clinical phenotypes.

Highlights

Mutations in HNRNPA1 encoding heterogeneous nuclear ribonucleoprotein A1 are a rare cause of amyotrophic lateral sclerosis (ALS) and multisystem proteinopathy (MSP). hnRNPA1 is part of the group of RNA-binding proteins (RBPs) that assemble with RNA to form RNPs. hnRNPs are concentrated in the nucleus and function in pre-mRNA splicing, mRNA stability, and the regulation of transcription and translation
Heterogeneous nuclear ribonucleoprotein A1 is a member of a large class of RNA-binding proteins (RBPs) that assemble with RNA to form ribonucleoproteins (RNPs). hnRNPs are evolutionarily conserved and are primarily localized to the nucleus, where they function in pre-mRNA splicing, mRNA stability, miRNA maturation, regulation of transcription and translation, and telomere biogenesis [1,2,3,4,5,6,7]
To screen for additional hnRNPA1 mutations in patients, we surveyed a large cohort of 547 patients with distal hereditary motor neuropathy (HMN), axonal Charcot-Marie-Tooth disease (CMT2), intermediate CMT (CMTi), spinal muscular atrophy (SMA), ALS, hereditary spastic paraplegia (HSP), and myopathy

Summary

Introduction

Mutations affecting several RBPs, including FUS, TDP-43, hnRNPA1, hnRNPA2B1, matrin-3, and TIA1, are linked to amyotrophic lateral sclerosis (ALS), frontotemporal dementia (FTD), multisystem proteinopathy (MSP), and hereditary inclusion body myopathy (hIBM) phenotypes [8, 9]. Missense mutations in the prion-like domain (PrLD) of hnRNPA1 (p.D262V/N and p.N267S) are linked to ALS (MIM 615426) and MSP3 (MIM 615424) [10, 11]. Additional ALS-linked mutations in hnRNPA1 are in the PrLD (p.G264R) [12] or the nuclear localization sequence (p.P288S/A) [13, 14]. Several modest-sized genetic screenings failed to identify further HNRNPA1 mutations in 2485 patients with ALS, FTD, MSP, hIBM, or other myopathies, suggesting that disease-linked mutations in HNRNPA1 may be rare [15,16,17,18,19]

Methods

Results

Conclusion