Human NK cell deficiency as a result of biallelic mutations in MCM10
Emily M. Mace, … , Anja K. Bielinsky, Jordan S. Orange
Emily M. Mace, … , Anja K. Bielinsky, Jordan S. Orange
Published August 31, 2020
Citation Information: J Clin Invest. 2020;130(10):5272-5286. https://doi.org/10.1172/JCI134966.
View: Text | PDF
Research Article Immunology

Human NK cell deficiency as a result of biallelic mutations in MCM10

Abstract

Human natural killer cell deficiency (NKD) arises from inborn errors of immunity that lead to impaired NK cell development, function, or both. Through the understanding of the biological perturbations in individuals with NKD, requirements for the generation of terminally mature functional innate effector cells can be elucidated. Here, we report a cause of NKD resulting from compound heterozygous mutations in minichromosomal maintenance complex member 10 (MCM10) that impaired NK cell maturation in a child with fatal susceptibility to CMV. MCM10 has not been previously associated with monogenic disease and plays a critical role in the activation and function of the eukaryotic DNA replisome. Through evaluation of patient primary fibroblasts, modeling patient mutations in fibroblast cell lines, and MCM10 knockdown in human NK cell lines, we have shown that loss of MCM10 function leads to impaired cell cycle progression and induction of DNA damage–response pathways. By modeling MCM10 deficiency in primary NK cell precursors, including patient-derived induced pluripotent stem cells, we further demonstrated that MCM10 is required for NK cell terminal maturation and acquisition of immunological system function. Together, these data define MCM10 as an NKD gene and provide biological insight into the requirement for the DNA replisome in human NK cell maturation and function.

Authors

Emily M. Mace, Silke Paust, Matilde I. Conte, Ryan M. Baxley, Megan M. Schmit, Sagar L. Patil, Nicole C. Guilz, Malini Mukherjee, Ashley E. Pezzi, Jolanta Chmielowiec, Swetha Tatineni, Ivan K. Chinn, Zeynep Coban Akdemir, Shalini N. Jhangiani, Donna M. Muzny, Asbjørg Stray-Pedersen, Rachel E. Bradley, Mo Moody, Philip P. Connor, Adrian G. Heaps, Colin Steward, Pinaki P. Banerjee, Richard A. Gibbs, Malgorzata Borowiak, James R. Lupski, Stephen Jolles, Anja K. Bielinsky, Jordan S. Orange

×

Figure 9

iPS cell–derived NK cells from the proband have impaired terminal maturation and increased replication stress.

Options: View larger image (or click on image) Download as PowerPoint
iPS cell–derived NK cells from the proband have impaired terminal matura...
iPS cells reprogrammed from patient primary fibroblasts were differentiated by teratoma formation to CD34+ HSCs, purified, and transplanted into NSG mice as described in Methods. Organs were harvested 21 days following transplantation, and human CD45+CD56+CD3− cells were analyzed for density of CD56 expression. n = 4 mice per genotype (patient and healthy donor–derived iPS cells). (A) Representative FACS histograms of NK cells from bone marrow of mice reconstituted with human NK cells from healthy donor– or patient-derived CD34+ cells generated from iPS cells. (B) Frequency of CD56bright NK cells from 4 mice per genotype from blood, spleen, and bone marrow, as indicated. NK cells were identified as human CD45+CD56+CD3− (bone marrow: 93–979 NK cells; blood: 20–405 NK cells; spleen: 60–880 NK cells, all from >106 cells from each organ per mouse), and the frequency of CD56bright NK cells based on CD56 density within the human NK cell population is shown. *P < 0.05, Mann-Whitney U test. (C) Splenocytes from mice transplanted with HD or patient-specific iPS cell–derived CD34+ cells were fixed, permeabilized, and incubated with anti-γH2AX antibody. Images were acquired by confocal microscopy. Scale bar: 5 μm. (D) Frequency of γH2AX foci per cell were enumerated by manual counting of 109 (HD) and 119 (patient) cells. ****P < 0.0001, Mann-Whitney U test. Data were pooled from 4 mice per genotype and are represented as mean ± 95% CI.