Preliminary Insights into RNA in CSF of Pediatric SMA Patients After 6 months of Nusinersen

Overview

Journal Biol Direct

Publisher Biomed Central

Specialty Biology

Date 2023 Sep 13

PMID 37705059

Authors

M Garofalo

S Bonanno

S Marcuzzo

C Pandini

E Scarian

F Dragoni

R Di Gerlando

M Bordoni

S Parravicini

C Gellera

R Masson

C Dosi

R Zanin

O Pansarasa

C Cereda

A Berardinelli

S Gagliardi

Affiliations

Soon will be listed here.

Abstract

Background: Spinal muscular atrophy (SMA) is a rare autosomal-recessive neurodegenerative disorder caused by mutations in survival motor neuron 1 (SMN1) gene, and consequent loss of function of SMN protein, which results in progressive loss of lower motor neurons, and muscular wasting. Antisense oligonucleotide (ASO) nusinersen (Spinraza®) modulates the pre-mRNA splicing of the SMN2 gene, allowing rebalance of biologically active SMN. It is administered intrathecally via lumbar puncture after removing an equal amount of cerebrospinal fluid (CSF). Its effect was proven beneficial and approved since 2017 for SMA treatment. Given the direct effect of nusinersen on RNA metabolism, the aim of this project was to evaluate cell-free RNA (cfRNA) in CSF of SMA patients under ASOs treatment for biomarker discovery.

Methods: By RNA-sequencing approach, RNA obtained from CSF of pediatric SMA type 2 and 3 patients was processed after 6 months of nusinersen treatment, at fifth intrathecal injection (T6), and compared to baseline (T0).

Results: We observed the deregulation of cfRNAs in patients at T6 and we were able to classify these RNAs into disease specific, treatment specific and treatment dependent. Moreover, we subdivided patients into "homogeneous" and "heterogeneous" according to their gene expression pattern. The "heterogeneous" group showed peculiar activation of genes coding for ribosomal components, meaning that in these patients a different molecular effect of nusinersen is observable, even if this specific molecular response was not referable to a clinical pattern.

Conclusions: This study provides preliminary insights into modulation of gene expression dependent on nusinersen treatment and lays the foundation for biomarkers discovery.

References

Lunn M, Wang C . Spinal muscular atrophy. Lancet. 2008; 371(9630):2120-33. DOI: 10.1016/S0140-6736(08)60921-6. View

Fang P, Li L, Zeng J, Zhou W, Wu W, Zhong Z . Molecular characterization and copy number of SMN1, SMN2 and NAIP in Chinese patients with spinal muscular atrophy and unrelated healthy controls. BMC Musculoskelet Disord. 2015; 16:11. PMC: 4328246. DOI: 10.1186/s12891-015-0457-x. View

Woo C, Maier V, Davey R, Brennan J, Li G, Brothers 2nd J . Gene activation of SMN by selective disruption of lncRNA-mediated recruitment of PRC2 for the treatment of spinal muscular atrophy. Proc Natl Acad Sci U S A. 2017; 114(8):E1509-E1518. PMC: 5338378. DOI: 10.1073/pnas.1616521114. View

Luchetti A, Ciafre S, Murdocca M, Malgieri A, Masotti A, Sanchez M . A Perturbed MicroRNA Expression Pattern Characterizes Embryonic Neural Stem Cells Derived from a Severe Mouse Model of Spinal Muscular Atrophy (SMA). Int J Mol Sci. 2015; 16(8):18312-27. PMC: 4581247. DOI: 10.3390/ijms160818312. View

Fallini C, Bassell G, Rossoll W . Spinal muscular atrophy: the role of SMN in axonal mRNA regulation. Brain Res. 2012; 1462:81-92. PMC: 3360984. DOI: 10.1016/j.brainres.2012.01.044. View

Finkel R, Mercuri E, Darras B, Connolly A, Kuntz N, Kirschner J . Nusinersen versus Sham Control in Infantile-Onset Spinal Muscular Atrophy. N Engl J Med. 2017; 377(18):1723-1732. DOI: 10.1056/NEJMoa1702752. View

Maggi L, Bello L, Bonanno S, Govoni A, Caponnetto C, Passamano L . Nusinersen safety and effects on motor function in adult spinal muscular atrophy type 2 and 3. J Neurol Neurosurg Psychiatry. 2020; 91(11):1166-1174. DOI: 10.1136/jnnp-2020-323822. View

Olsson B, Alberg L, Cullen N, Michael E, Wahlgren L, Kroksmark A . NFL is a marker of treatment response in children with SMA treated with nusinersen. J Neurol. 2019; 266(9):2129-2136. PMC: 6687695. DOI: 10.1007/s00415-019-09389-8. View

Fatima R, Akhade V, Pal D, Rao S . Long noncoding RNAs in development and cancer: potential biomarkers and therapeutic targets. Mol Cell Ther. 2015; 3:5. PMC: 4469312. View

10.

Wang J, Chen J, Sen S . MicroRNA as Biomarkers and Diagnostics. J Cell Physiol. 2015; 231(1):25-30. PMC: 8776330. DOI: 10.1002/jcp.25056. View

11.

Podda M, Bacciu D, Micheli A, Bellu R, Placidi G, Gagliardi L . A machine learning approach to estimating preterm infants survival: development of the Preterm Infants Survival Assessment (PISA) predictor. Sci Rep. 2018; 8(1):13743. PMC: 6137213. DOI: 10.1038/s41598-018-31920-6. View

12.

Teunissen C, Verheul C, Willemse E . The use of cerebrospinal fluid in biomarker studies. Handb Clin Neurol. 2017; 146:3-20. DOI: 10.1016/B978-0-12-804279-3.00001-0. View

13.

Bonanno S, Marcuzzo S, Malacarne C, Giagnorio E, Masson R, Zanin R . Circulating MyomiRs as Potential Biomarkers to Monitor Response to Nusinersen in Pediatric SMA Patients. Biomedicines. 2020; 8(2). PMC: 7168147. DOI: 10.3390/biomedicines8020021. View

14.

Catapano F, Zaharieva I, Scoto M, Marrosu E, Morgan J, Muntoni F . Altered Levels of MicroRNA-9, -206, and -132 in Spinal Muscular Atrophy and Their Response to Antisense Oligonucleotide Therapy. Mol Ther Nucleic Acids. 2016; 5(7):e331. PMC: 5014531. DOI: 10.1038/mtna.2016.47. View

15.

Riitano G, Capozzi A, Recalchi S, Caissutti D, Longo A, Mattei V . Anti-β2-GPI Antibodies Induce Endothelial Cell Expression of Tissue Factor by LRP6 Signal Transduction Pathway Involving Lipid Rafts. Cells. 2022; 11(8). PMC: 9025633. DOI: 10.3390/cells11081288. View

16.

Subramanian A, Tamayo P, Mootha V, Mukherjee S, Ebert B, Gillette M . Gene set enrichment analysis: a knowledge-based approach for interpreting genome-wide expression profiles. Proc Natl Acad Sci U S A. 2005; 102(43):15545-50. PMC: 1239896. DOI: 10.1073/pnas.0506580102. View

17.

Hall A, Lalli G . Rho and Ras GTPases in axon growth, guidance, and branching. Cold Spring Harb Perspect Biol. 2010; 2(2):a001818. PMC: 2828272. DOI: 10.1101/cshperspect.a001818. View

18.

Spillane M, Gallo G . Involvement of Rho-family GTPases in axon branching. Small GTPases. 2014; 5:e27974. PMC: 4114606. DOI: 10.4161/sgtp.27974. View

19.

Zheleznyakova G, Voisin S, Kiselev A, Almen M, Xavier M, Maretina M . Genome-wide analysis shows association of epigenetic changes in regulators of Rab and Rho GTPases with spinal muscular atrophy severity. Eur J Hum Genet. 2013; 21(9):988-93. PMC: 3746269. DOI: 10.1038/ejhg.2012.293. View

20.

Rubio-Gozalbo M, Smeitink J, Ruitenbeek W, Ter Laak H, Mullaart R, Schuelke M . Spinal muscular atrophy-like picture, cardiomyopathy, and cytochrome c oxidase deficiency. Neurology. 1999; 52(2):383-6. DOI: 10.1212/wnl.52.2.383. View