Chromosomal translocation t(11;14) and p53 deletion induced by the CRISPR/Cas9 system in normal B cell-derived iPS cells

• Published in: Scientific reports Vol. 11; no. 1; pp. 5216 - 13
• Main Authors: Azami, Yusuke, Tsuyama, Naohiro, Abe, Yu, Sugai-Takahashi, Misaki, Kudo, Ken-ichi, Ota, Akinobu, Sivasundaram, Karnan, Muramatsu, Moe, Shigemura, Tomonari, Sasatani, Megumi, Hashimoto, Yuko, Saji, Shigehira, Kamiya, Kenji, Hanamura, Ichiro, Ikezoe, Takayuki, Onodera, Masafumi, Sakai, Akira
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 04.03.2021; Nature Publishing Group; Nature Portfolio
• Subjects: 631/532; 631/80; Alleles; B-Lymphocytes - metabolism; B-Lymphocytes - pathology; Cell culture; Cell differentiation; Cell Differentiation - genetics; Cell Line, Tumor; Chromosome Aberrations; Chromosome deletion; Chromosomes, Human, Pair 11 - genetics; Chromosomes, Human, Pair 14 - genetics; Cord blood; CRISPR; CRISPR-Cas Systems - genetics; Cyclin D1 - genetics; Heavy chains; Hematopoietic Stem Cells; Humanities and Social Sciences; Humans; Immunoglobulin Heavy Chains - genetics; Immunoglobulins; In Situ Hybridization, Fluorescence; Induced Pluripotent Stem Cells - metabolism; Induced Pluripotent Stem Cells - pathology; Lymphocytes; Lymphocytes B; multidisciplinary; Multiple myeloma; Multiple Myeloma - genetics; p53 Protein; Pluripotency; Progenitor cells; Science; Science (multidisciplinary); Stem cells; Translocation; Translocation, Genetic - genetics; Tumor Suppressor Protein p53 - genetics; VDJ Exons - genetics
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-021-84628-5

Multiple myeloma (MM) cells are derived from mature B cells based on immunoglobulin heavy chain ( IgH ) gene analysis. The onset of MM is often caused by a reciprocal chromosomal translocation (cTr) between chr 14 with IgH and chr 11 with CCND1 . We propose that mature B cells gain potential to transform by reprograming, and then chromosomal aberrations cause the development of abnormal B cells as a myeloma-initiating cell during B cell redifferentiation. To study myeloma-initiating cells, we have already established normal B cell-derived induced pluripotent stem cells (BiPSCs). Here we established two BiPSCs with reciprocal cTr t(11;14) using the CRISPR/Cas9 system; the cleavage site were located in the IgH Eμ region of either the VDJ rearranged allele or non-rearranged allele of IgH and the 5′-upsteam region of the CCND1 (two types of BiPSC13 with t(11;14) and MIB2-6 with t(11;14)). Furthermore, p53 was deleted using the CRISPR/Cas9 system in BiPSC13 with t(11;14). These BiPSCs differentiated into hematopoietic progenitor cells (HPCs). However, unlike cord blood, those HPCs did not differentiated into B lymphocytes by co-culture with BM stromal cell. Therefore, further ingenuity is required to differentiate those BiPSCs-derived HPCs into B lymphocytes.