Development of a gene therapy for congenital phagocyte disorder using both genome editing and iPS cell technology

2017 Fiscal Year Final Research Report

• Project/Area Number: 15H04874
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Pediatrics
• Research Institution: Shinshu University
• Principal Investigator: Koike Kenichi 信州大学, 医学部, 特任教授 (40143979)
• Co-Investigator(Kenkyū-buntansha): 中沢 洋三 信州大学, 医学部, 教授 (60397312) ; 重村 倫成 信州大学, 学術研究院医学系(医学部附属病院), 講師 (70623916)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Keywords: iPS細胞 / ゲノム修飾 / 先天性好中球減少症 / 遺伝子治療

Outline of Final Research Achievements

In parallel with genetic editing experiments of iPS cells established from a patient with ELANE-mutated severe congenital neutropenia, we performed genetic editing experiments on iPS cells derived from a patient with juvenile myelomonocytic leukemia (JMML). Both iPS cells with PTPN11 gene mutation and those with wild type of PTPN11gene were established from a JMML patient. Mutant iPS cells possessed significantly higher ability to generate CD34 positive cells. In addition, whole-exome sequencing and Sanger sequencing revealed that JMML iPSCs with PTPN11 mutation possessed OSBP2 mutation, while JMML iPSCs with no PTPN11 mutation did not carry OSBP2 mutation. By homologous recombination using zinc finger nuclease, we corrected PTPN11 mutation in iPSCs with both PTPN11 mutation and OSBP2 mutation, and transduced PTPN11 mutation into JMML iPSCs with wild type of the two genes. The results suggest that PTPN11 mutation itself contributes to leukemogenesis of this patient.

Free Research Field

小児科学（血液腫瘍学）