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Abstract: TH-PO537

Expression of GLA, HNRNPH2, and RPL36A Genes in Fabry Disease Male and Female Cell Lines

Session Information

  • Pathology and Lab Medicine
    November 03, 2022 | Location: Exhibit Hall, Orange County Convention Center‚ West Building
    Abstract Time: 10:00 AM - 12:00 PM

Category: Pathology and Lab Medicine

  • 1700 Pathology and Lab Medicine

Authors

  • Islam, Md Saimul, Texas Tech Health Science Center, Amarillo, Texas, United States
  • Al-Obaide, Mohammed A., Texas Tech Health Science Center, Amarillo, Texas, United States
  • Vasylyeva, Tetyana L., Texas Tech Health Science Center, Amarillo, Texas, United States
Background

Fabry disease (FD) is a rare genetic disorder caused by mutations in the α-galactosidase A (GLA) gene resulting in α-galactosidase An enzyme deficiency. Despite, enzyme replacement therapy, FD patients often remain symptomatic indicating that other mechanisms might be associated with this disease. Our previous in-vitro study showed that knockdown of the first locus (RPL36A) in the readthrough transcription RPL36A-HNRNPH2 locus leads to decreased expression of GLA, RPL36A, and HNRNPH2. We also observed GLA c.1033_1034delTC deletion in male FD patients greatly reduces the expression levels of GLA, RPL36A, and HNRNPH2, and at lower, but significant levels in females, as compared with healthy males and females. Based on this finding we have hypothesized that regulation of GLA genes by its adjacent loci RPL36A and HNRNPH2 might play a significant role during the pathogenesis of FD. To analyze the expression status of GLA, RPL36A and HNRNPH2 genes in different male and female FD patient-derived cell lines.

Methods

FD cell lines were purchased for three female and male patients along with one fibroblast-derived control cell line from Coriell Institute for Medical Research, and were grown as per supplier instructions. Then, harvested cell mRNA expression analysis by qRT-PCR was performed.

Results

qRT-PCR revealed that GLA mRNA expression was downregulated in both female and male FD cell lines except for Gm0881 (FD MALE with R220X) Unexpectedly, GLA had more reduction in expression in female than male cell lines. Although the Gm0881 cell line showed high GLA mRNA expression, it has no GLA enzymatic activity due to c.658C>T in exon 5, resulting in a stop codon [Arg220Ter (R220X)]. This suggested the key functional importance of this mutation in GLA activity vs. GLA expression. Downregulation of HNRNPH2 expression was also observed in the FD cell lines irrespective of male and female source, except in the Gm02774 cell line. But, unlike GLA expression, HNRNPH2 had a higher reduction in the male than the female cell lines. Similarly. RPL36A was downregulated in all FD cell lines irrespective of whether the cells were male or female-derived.

Conclusion

Dysregulation of expression of GLA and its two conjugative loci RPL36A and HNRNPH2 demonstrate differences based on sex in cell lines derived from male and female FD patients.

Funding

  • Commercial Support