Establishment of iPSC lines and zebrafish with loss-of-function AHDC1 variants: Models for Xia-Gibbs syndrome

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Carvalho, Laura Machado Lara ; Branco, Elisa Varella ; Sarafian, Raquel Delgado ; Kobayashi, Gerson Shigeru ; de Araujo, Fabiano Tofoli ; Souza, Lucas Santos ; Moreira, Danielle de Paula ; Hsia, Gabriella Shih Ping ; Bertollo, Eny Maria Goloni ; Buck, Cecilia Barbosa ; da Costa, Silvia Souza ; Fialho, Davi Mendes ; Vasconcelos, Felipe Tadeu Galante Rocha de ; Brito, Luciano Abreu ; Machado, Luciana Elena de Souza Fraga ; Ramos, Igor Cabreira ; Pereira, Lygia da Veiga ; Koiffmann, Celia Priszkulnik ; Passos-Buen, Maria Rita dos Santos e ; Mendes, Tiago Antonio de Oliveira ; Krepischi, Ana Cristina Victorino ; Rosenberg, Carla

• Document type: Journal article
• Source: Gene; v. 871, p. 10-pg., 2023-04-22.

Abstract

Xia-Gibbs syndrome (XGS) is a syndromic form of intellectual disability caused by heterozygous AHDC1 variants, but the pathophysiological mechanisms underlying this syndrome are still unclear. In this manuscript, we describe the development of two different functional models: three induced pluripotent stem cell (iPSC) lines with different loss-of-function (LoF) AHDC1 variants, derived by reprogramming peripheral blood mononuclear cells from XGS patients, and a zebrafish strain with a LoF variant in the ortholog gene (ahdc1) obtained through CRISPR/Cas9-mediated editing. The three iPSC lines showed expression of pluripotency factors (SOX2, SSEA-4, OCT3/4, and NANOG). To verify the capacity of iPSC to differentiate into the three germ layers, we obtained embryoid bodies (EBs), induced their differentiation, and confirmed the mRNA expression of ectodermal, mesodermal, and endodermal markers using the TaqMan hPSC Scorecard. The iPSC lines were also approved for the following quality tests: chromosomal microarray analysis (CMA), mycoplasma testing, and short tandem repeat (STR) DNA profiling. The zebrafish model has an insertion of four base pairs in the ahdc1 gene, is fertile, and breeding between heterozygous and wild-type (WT) animals generated offspring in a genotypic proportion in agreement with Mendelian law. The established iPSC and zebrafish lines were deposited on the hpscreg.eu and zfin.org platforms, respectively. These biological models are the first for XGS and will be used in future studies that investigate the pathophysiology of this syndrome, unraveling its underlying molecular mechanisms. (AU)

• FAPESP's process: 20/03108-0 - Establishment of the in vitro cardiovascular model derived from human Pluripotent Stem Cells (hiPSCs) for functional analysis of Hypertension
• Grantee: Raquel Delgado Sarafian
• Support Opportunities: Scholarships in Brazil - Doctorate
• FAPESP's process: 20/04744-8 - Evaluation of the muscle regeneration process in limb-girdle Muscular Dystrophy 2B
• Grantee: Lucas Santos e Souza
• Support Opportunities: Scholarships in Brazil - Doctorate
• FAPESP's process: 13/08028-1 - CEGH-CEL - Human Genome and Stem Cell Research Center
• Grantee: Mayana Zatz
• Support Opportunities: Research Grants - Research, Innovation and Dissemination Centers - RIDC
• FAPESP's process: 20/10168-0 - Structural and functional characterization of mitochondrial protein phosphatases by redox metabolism and their role in cell biology
• Grantee: Luciana Elena de Souza Fraga Machado
• Support Opportunities: Scholarships in Brazil - Young Researchers
• FAPESP's process: 18/08486-3 - Genetic bases of Syndromic Obesity and molecular mechanisms involved in its physiopathology
• Grantee: Laura Machado Lara Carvalho
• Support Opportunities: Scholarships in Brazil - Doctorate
• FAPESP's process: 19/02605-3 - Structural and functional characterization of mitochondrial protein phosphatases by redox metabolism and their role in cell biology
• Grantee: Luciana Elena de Souza Fraga Machado
• Support Opportunities: Research Grants - Young Investigators Grants