Development of methods for genetic analysis of malignant lymphoma using cell free DNA from peripheral blood and FFPE DNA

• Project/Area Number: 26893107
• Research Category: Grant-in-Aid for Research Activity Start-up
• Allocation Type: Single-year Grants
• Research Field: Hematology
• Research Institution: Nagoya University
• Principal Investigator: IRIYAMA Chisako 名古屋大学, 医学部附属病院, 病院助教 (40732631)
• Project Period (FY): 2014-08-29 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥2,990,000 (Direct Cost: ¥2,300,000、Indirect Cost: ¥690,000); Fiscal Year 2015: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2014: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
• Keywords: 悪性リンパ腫 / 末梢血無細胞遊離DNA / FFPE DNA / 遺伝子変異解析 / 末梢血遊離DNA / 遺伝子解析

Outline of Final Research Achievements

Analysis were performed using cell free DNA (cfDNA) from peripheral blood and DNA from formalin fixed paraffin embedded tissue (FFPE DNA) of patients with diffuse large B cell lymphoma (DLBCL). Concentration of cfDNA varied among patients (0.23 to 547 ng in 1ml plasma), and changed during chemotherapy. cfDNA was detected as ladder pattern. Amplification of 300 to 600 base pairs of PCR products was successfully performed using cfDNA. Genomic mutations which were detected in biopsy specimen can be detected in cfDNA. It is suggested that cfDNA can be alternative source for genetic analysis which can be obtained less invasively and frequently in order to detect genetic alteration time-dependently. FFPE DNA was detected as smear pattern and sometimes it was difficult to amplify DNA with PCR for some samples. However it enables us to analyze genetic abnormalities retrospectively for many cases whose fresh biopsy samples could not be acquired enough.