A Mouse Model with Ablated Asparaginase and Isoaspartyl Peptidase 1 () Develops Early Onset Retinal Degeneration (RD) Recapitulating the Human Phenotype

Overview

Journal Genes (Basel)

Publisher MDPI

Date 2022 Aug 26

PMID 36011372

Authors

Pooja Biswas

Anne Marie Berry

Qais Zawaydeh

Dirk-Uwe G Bartsch

Pongali B Raghavendra

J Fielding Hejtmancik

Naheed W Khan

S Amer Riazuddin

Radha Ayyagari

Affiliations

Soon will be listed here.

Abstract

We previously identified a homozygous G178R mutation in human () through whole-exome analysis responsible for early onset retinal degeneration (RD) in patients with cone-rod dystrophy. The mutant G178R ASRGL1 expressed in Cos-7 cells showed altered localization, while the mutant ASRGL1 in lacked the autocatalytic activity needed to generate the active protein. To evaluate the effect of impaired ASRGL1 function on the retina in vivo, we generated a mouse model with c.578_579insAGAAA (NM_001083926.2) mutation () through the CRISPR/Cas9 methodology. The expression of ASGRL1 and its asparaginase activity were undetectable in the retina of mice. The ophthalmic evaluation of mice showed a significant and progressive decrease in scotopic electroretinographic (ERG) response observed at an early age of 3 months followed by a decrease in photopic response around 5 months compared with age-matched wildtype mice. Immunostaining and RT-PCR analyses with rod and cone cell markers revealed a loss of cone outer segments and a significant decrease in the expression of , , and at 3 months in mice compared with age-matched wildtype mice. Importantly, the retinal phenotype of mice is consistent with the phenotype observed in patients harboring the G178R mutation in confirming a critical role of ASRGL1 in the retina and the contribution of mutations in retinal degeneration.

References

Lev S . Molecular aspects of retinal degenerative diseases. Cell Mol Neurobiol. 2002; 21(6):575-89. PMC: 11533866. DOI: 10.1023/a:1015183500719. View

Daiger S, Sullivan L, Bowne S . Genes and mutations causing retinitis pigmentosa. Clin Genet. 2013; 84(2):132-41. PMC: 3856531. DOI: 10.1111/cge.12203. View

Collin G, Gogna N, Chang B, Damkham N, Pinkney J, Hyde L . Mouse Models of Inherited Retinal Degeneration with Photoreceptor Cell Loss. Cells. 2020; 9(4). PMC: 7227028. DOI: 10.3390/cells9040931. View

Zhou Y, Tian W, Jiang X, Yang H, Jiang Z, Li X . Deletion of Leads to Photoreceptor Degeneration in Mice. Front Cell Dev Biol. 2022; 9:783547. PMC: 8805730. DOI: 10.3389/fcell.2021.783547. View

Chavali V, Khan N, Cukras C, Bartsch D, Jablonski M, Ayyagari R . A CTRP5 gene S163R mutation knock-in mouse model for late-onset retinal degeneration. Hum Mol Genet. 2011; 20(10):2000-14. PMC: 3080610. DOI: 10.1093/hmg/ddr080. View

Petersen-Jones S . Animal models of human retinal dystrophies. Eye (Lond). 1998; 12 ( Pt 3b):566-70. DOI: 10.1038/eye.1998.146. View

Biswas P, Chavali V, Agnello G, Stone E, Chakarova C, Duncan J . A missense mutation in ASRGL1 is involved in causing autosomal recessive retinal degeneration. Hum Mol Genet. 2016; 25(12):2483-2497. PMC: 6086560. DOI: 10.1093/hmg/ddw113. View

Fauser S, Luberichs J, Schuttauf F . Genetic animal models for retinal degeneration. Surv Ophthalmol. 2002; 47(4):357-67. DOI: 10.1016/s0039-6257(02)00314-4. View

Radadiya A, Zhu W, Coricello A, Alcaro S, Richards N . Improving the Treatment of Acute Lymphoblastic Leukemia. Biochemistry. 2020; 59(35):3193-3200. PMC: 7497903. DOI: 10.1021/acs.biochem.0c00354. View

10.

Li W, Cantor J, Yogesha S, Yang S, Chantranupong L, Liu J . Uncoupling intramolecular processing and substrate hydrolysis in the N-terminal nucleophile hydrolase hASRGL1 by circular permutation. ACS Chem Biol. 2012; 7(11):1840-7. PMC: 3514461. DOI: 10.1021/cb300232n. View

11.

Kumar K, Kaur J, Walia S, Pathak T, Aggarwal D . L-asparaginase: an effective agent in the treatment of acute lymphoblastic leukemia. Leuk Lymphoma. 2013; 55(2):256-62. DOI: 10.3109/10428194.2013.803224. View

12.

Inui M, Miyado M, Igarashi M, Tamano M, Kubo A, Yamashita S . Rapid generation of mouse models with defined point mutations by the CRISPR/Cas9 system. Sci Rep. 2014; 4:5396. PMC: 4066261. DOI: 10.1038/srep05396. View

13.

Cantor J, Stone E, Chantranupong L, Georgiou G . The human asparaginase-like protein 1 hASRGL1 is an Ntn hydrolase with beta-aspartyl peptidase activity. Biochemistry. 2009; 48(46):11026-31. PMC: 2782781. DOI: 10.1021/bi901397h. View

14.

Ferrari S, Di Iorio E, Barbaro V, Ponzin D, Sorrentino F, Parmeggiani F . Retinitis pigmentosa: genes and disease mechanisms. Curr Genomics. 2011; 12(4):238-49. PMC: 3131731. DOI: 10.2174/138920211795860107. View

15.

Li W, Irani S, Crutchfield A, Hodge K, Matthews W, Patel P . Intramolecular Cleavage of the hASRGL1 Homodimer Occurs in Two Stages. Biochemistry. 2016; 55(6):960-9. PMC: 4809669. DOI: 10.1021/acs.biochem.5b01157. View

16.

Wang H, Yang H, Shivalila C, Dawlaty M, Cheng A, Zhang F . One-step generation of mice carrying mutations in multiple genes by CRISPR/Cas-mediated genome engineering. Cell. 2013; 153(4):910-8. PMC: 3969854. DOI: 10.1016/j.cell.2013.04.025. View

17.

Qin W, Kutny P, Maser R, Dion S, Lamont J, Zhang Y . Generating Mouse Models Using CRISPR-Cas9-Mediated Genome Editing. Curr Protoc Mouse Biol. 2016; 6(1):39-66. PMC: 4848752. DOI: 10.1002/9780470942390.mo150178. View

18.

Mandal M, Vasireddy V, Jablonski M, Wang X, Heckenlively J, Hughes B . Spatial and temporal expression of MFRP and its interaction with CTRP5. Invest Ophthalmol Vis Sci. 2006; 47(12):5514-21. DOI: 10.1167/iovs.06-0449. View

19.

Hamel C . Retinitis pigmentosa. Orphanet J Rare Dis. 2006; 1:40. PMC: 1621055. DOI: 10.1186/1750-1172-1-40. View

20.

Edqvist P, Huvila J, Forsstrom B, Talve L, Carpen O, Salvesen H . Loss of ASRGL1 expression is an independent biomarker for disease-specific survival in endometrioid endometrial carcinoma. Gynecol Oncol. 2015; 137(3):529-37. DOI: 10.1016/j.ygyno.2015.03.055. View