Novel diagnostic approaches for Fanconi anemia (FA) by single-cell sequencing and capillary nano-immunoassay

Abstract

Next-generation sequencing technology has been widely utilized for the diagnosis of Fanconi anemia (FA). However, mixed cell sequencing and chimerism of FA patients may lead to unconfirmed genetic subtypes. Herein, we introduced two novel diagnostic methods, including single-cell sequencing and capillary nano-immunoassay. One FA case with FANCM c.4931G>A p.R1644Q and FANCD1 c.6325G>A p.V2109I was studied. The DNA of 28 cells was amplified and eight types of cells were observed after Sanger sequencing. There were two homozygous mutations ( FANCM / FANCD1 ). Furthermore, the capillary nano-immunoassay was conducted to analyze the expression profile of FA-associated proteins. Abnormal FANCM and FANCD1 expressions simultaneously existed. This case was thus diagnosed as FA-D1/FA-M dual subtype. Compared with mixed cell sequencing, single-cell sequencing data shows more accuracy for the FA subtype evaluation, while the capillary nano-immunoassay is a good method to detect the expression profile of abnormal or modified FA protein.

DNAH2 facilitates the homologous recombination repair of Fanconi anemia pathway through modulating FANCD2 ubiquitination

“…Fanconi anemia (FA) is characterized by progressive bone marrow failure, congenital abnormalities, and cancer susceptibility 1–5 . Cells derived from FA patients display distinct patterns of chromosomal abnormalities 1–6 . In addition, these cells are hypersensitive to DNA crosslinking agents, such as mitomycin C (MMC) 7,8 .…”

“…[1][2][3][4][5] Cells derived from FA patients display distinct patterns of chromosomal abnormalities. [1][2][3][4][5][6] In addition, these cells are hypersensitive to DNA crosslinking agents, such as mitomycin C (MMC). 7,8 There are more than 20 genes identified in the DNA repair pathway of FA.…”