Fig 2: KAT6B mutations in HEK293T cells result in dysregulation of relevant gene regulatory pathways. (a) For each of the KAT6B‐mutant cell lines generated by Cas9‐CRISPR, we sequenced genomic DNA and cDNA to identify allelic mutations in each line. The mutations are predicted to result in frameshift mutations resulting in protein truncation. Mut1 cell line has a single mutation, whereas Mut2 has two independent mutations on three alleles. (b) Heat map comparing counts for all differentially expressed genes with fold‐change >1.5 and FDR <0.5 (n = 434 genes). We compared our HEK293 control cells (C1 and C2) and the cells with KAT6B mutation (Mut1 and Mut2). (c) Differentially expressed genes were found to be significantly enriched in core features related to KAT6B clinical syndrome including skeletal ossification, urogenital development, axonal development. The X‐axis demonstrates the number of differentially expressed genes that are within each category on the Y‐axis. The color of the bar represents the adjusted p‐value for the enrichment

Fig 3: Genetic Counseling support related to the care and management of patients. (a) Of the 18 respondents, a genetic counselor was present for 0–4 sessions involving the team involved in the care of the patient. (b) Individuals received a diagnosis of a KAT6B-related disorder as early as 6 weeks old to as late as 23 years old. (c) Typical roles and responsibilities that genetic counselors hold demonstrate consistency despite the heterogeneous nature of patients with mutations in KAT6B. (d) Overall, patients and their families were receptive to the information given at diagnosis and relieved to have a diagnosis. (e) Many families sought additional information about the likely or expected development of disease with extra concern for future medical complications. (f) Due to the phenotypic heterogeneity found among patients with pathogenic variants in KAT6B, a wide variety of challenges arose when relaying information about the condition to the family

Fig 4: Pathogenic variants in KAT6B. We added 10 novel variants from our cohort to the list of previously reported variants. The novel pathogenic variants in our cohort are shown above the gene; previously reported variants are displayed below the gene. Genitopatellar (GPS)-related variants are denoted in orange, Say-Barber-Biesecker-Young-Simpson (SBBYS)-related variants are denoted in blue, and intermediate phenotype-related variants are denoted in black. RefSeq ID for KAT6B is NM_012330.3. Various protein domains are NEMM domain (AA 1–176), PHD domains (AA 177–360), HAT domain (AA 361–1070), acidic domain (AA 1071–1417), and Ser/Met domain (AA 1418–2073)

Fig 5: Lysine acetyltransferase 6B (KAT6B)‐dependent nuclear factor‐kappa B (NF‐κB) signalling is responsible for living tumour cell repopulation stimulated by dying cells. (A) Briefly, SCC‐25, SCC‐15 and SCC‐9 cells were treated with cisplatin (cDDP) for 24 h. Then, another SCC‐25, SCC‐15 and SCC‐9 cells were seeded among each cDDP‐treated cells. The transactivation of NF‐κB signalling in cells was measured by Cignal Reporter Assays (n = 3), *** p < .001. (B) SCC‐25, SCC‐15 and SCC‐9 cells were treated with cDDP for 24 h. Then, new SCC‐25, SCC‐15 and SCC‐9 cells were seeded among each cDDP‐treated cells. The protein levels of IκBα, p‐IκBα, CCND1, Bcl2 and β‐Actin in these cells were detected by western blots. (C) SCC‐25, SCC‐15 and SCC‐9 feeder cells were treated with cDDP for 24 h. SCC‐25/Luc, SCC‐15/Luc and SCC‐9/Luc reporter cells were transfected with pBabe‐Con or pBabe‐IκBα, and then seeded among the respective feeder cells with cDDP treatment or alone in 24‐well plates. Cancer cell repopulation in vitro was evaluated by luciferase activities, ** p < .01. (D) SCC‐25 cells were treated with cDDP for 24 h. SCC‐25/Luc reporter cells were transfected with pBabe‐Con or pBabe‐IκBα and injected subcutaneously together with cDDP‐treated SCC‐25 cells into nude mice. The growth of SCC‐25/Luc cells transfected with pBabe‐Con or pBabe‐IκBα was represented by luciferase levels and tumor size, ** p < .01, *** p < .001. (E) SCC‐25, SCC‐15 and SCC‐9 cells were cocultured with cDDP‐treated each feeder cells. The protein levels of KAT6B, H3K9Ac, H3K14Ac and β‐Actin were detected by western blots. (F and G) SCC‐25 cells were treated with cDDP for 24 h, another SCC‐25 cells were transfected with shRNAs specifically targeting KAT6B and then cocultured with cDDP‐treated SCC‐25 cells. The expression levels of CCND1 and Bcl2 were detected by quantitative real‐time polymerase chain reaction (qRT‐PCR) (n = 3) (F). The H3K9Ac levels at the promoter regions of CCND1 and Bcl2 were detected by chromatin immunoprecipitation (ChIP)‐qPCR (n = 3), ** p < .01, *** p < .001 (G). (H) SCC‐25 cells were treated with cDDP for 24 h, another SCC‐25 cells were cocultured with cDDP‐treated SCC‐25 cells. Total proteins were subjected to immunoprecipitation (IP) using an anti‐p65 antibody or control immunoglobulin G (IgG), followed by western blot analysis with a specific antibody against KAT6B. (I) SCC‐25 cells were treated with cDDP for 24 h. SCC‐25/Luc, SCC‐15/Luc and SCC‐9/Luc reporter cells were transfected with shRNAs specifically targeting KAT6B and then seeded among the respective feeder cells with cDDP treatment or alone in 24‐well plates. Cancer cell repopulation in vitro was observed by luciferase activities, ** p < .01. (J) SCC‐25 cells were treated with cDDP for 24 h. SCC‐25/Luc cells with or without KAT6B knockdown were injected subcutaneously together with cDDP‐treated SCC‐25 cells in nude mice, and tumour growth is represented by luciferase levels, ** p < .01, *** p < .001. Data in (A), (C), (D), (F), (G), (I) and (J) are represented as the mean ±SEM. Statistical significance was determined by a two‐tailed Student's t‐test