A novel monogenic preimplantation genetic testing strategy for sporadic polycystic kidney caused by de novoPKD1 mutation

Abstract

Autosomal dominant hereditary polycystic kidney disease (ADPKD) is the most common inherited kidney disease that causes end‐stage renal disease and kidney failure. Preimplantation genetic testing for monogenic (PGT‐M) can effectively prevent the transmission of genetic diseases from parents to the offspring before pregnancy. However, PGT‐M currently adopts the single nucleotide polymorphism (SNP) linkage analysis for embryo's pathogenic gene carrying status and linkage analysis requires proband of the family. Here we report a new PGT‐M strategy using single sperm SNP linkage analysis for male patient with sporadic ADPKD caused by de novo PKD1 mutation. We recruited five couples with male patient with ADPKD caused by de novo PKD1 mutation, and 39 embryos from six PGT‐M cycles were detected. The five couples had at least one embryo that does not carry the PKD1 mutation. Within these five couples, the accuracy of carrier status of embryos was confirmed by amniotic fluid gene detection of two couples and two couples successfully delivered healthy fetuses. Therefore, the new PGT‐M strategy of using single sperm SNP linkage analysis was proved to be feasible and effective for male patient with ADPKD caused by de novo PKD1 mutation.

A long-read sequencing and SNP haplotype-based novel preimplantation genetic testing method for female ADPKD patient with de novo PKD1 mutation

“…However, for approximately 10–15% of the ADPKD patients with de novo mutations in PKD1 or without a proband, haplotype analysis is challenging. For male ADPKD patients with de novo PKD1 mutations, an effective haplotype strategy based on single sperm was carried out to analyze the carrying status of embryos [ 27 ]. But for female patients, the gametes or PB cells are relatively hard to obtain, which makes haplotype analysis even harder.…”

Combined Preimplantation Genetic Testing for Genetic Kidney Disease: Genetic Risk Identification, Assisted Reproductive Cycle, and Pregnancy Outcome Analysis

“…However, it is not allowed in China for ethical reasons and some patients who came to our IVF center for consultation would choose to reduce the risk through egg donation. Furthermore, for special cases, including de novo (31) and germline mosaicism (32) pathogenic variants in husband or wife, PGT-M currently adopts the strategy of constructing haploid for linkage analysis by SNPs through nextgeneration sequencing (NGS) or Karyomapping array combined with direct sequencing methods such as Sanger sequencing. Our center has carried out relevant cases and achieved successful healthy live births.…”

State of the Science and Ethical Considerations for Preimplantation Genetic Testing for Monogenic Cystic Kidney Diseases and Ciliopathies

“…However, family members are often not available, and linkage analysis cannot be performed with de novo variants and instead need to be identified in single gametes. While single gamete (polar body or sperm) SNP linkage analysis for patients with de novo PKD1 pathogenic variants has been reported, 55,56 it poses a significant challenge due to its multiple duplicated genomic regions. Successful PGT-M for a pseudogene-homologous variant in PKD1 has been reported through a whole genome amplification approach using single sperm or blastomeres, followed by haplotype construction and analysis through long-range PCR has been reported.…”

“…50 Challenges and Technical Considerations of PGT-M for ADPKD Some technical limitations exist for PGT-M, with ADPKD presenting unique challenges. PKD1 lies in a complex, segmentally duplicated region with six pseudogenes (Figure 5), making it a challenging target and necessitating unique methodology for PGT-M. [51][52][53][54][55][56] Karyomapping (Figure 2) with flanking SNPs could potentially be used to overcome these challenges. However, a minimum of two affected family members with a clear phenotype and ideally from two different generations is usually required.…”