TCR repertoire analysis targeting HTLV-1-infected cells in HAM patients for biomarker discovery

• Project/Area Number: 20K22879
• Research Category: Grant-in-Aid for Research Activity Start-up
• Allocation Type: Multi-year Fund
• Review Section: 0902:General internal medicine and related fields
• Research Institution: Kagoshima University
• Principal Investigator: Satoshi Nozuma 鹿児島大学, 医歯学域医学系, 助教 (40884052)
• Project Period (FY): 2020-09-11 – 2022-03-31
• Project Status: Completed (Fiscal Year 2021)
• Budget Amount: ¥2,860,000 (Direct Cost: ¥2,200,000、Indirect Cost: ¥660,000); Fiscal Year 2021: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2020: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
• Keywords: HTLV-1関連脊髄症 / 神経免疫性疾患 / 神経感染症 / T細胞受容体レパトア解析 / HTLV-1 / T細胞受容体レパトア / HAM / 神経免疫疾患

Outline of Research at the Start

HAMはHTLV-1ウイルスに感染したTリンパ球の増殖とそれに対する特異的な免疫反応が病態の中心である。近年TCRレパトア研究が癌、免疫、感染症領域で進められ、細胞性免疫の評価やウイルス抗原特異的T細胞の同定に利用されている。本研究では感染細胞を標的にHAMに特異的なTCRクローン型を同定することで、診断のバイオマーカーとなることが期待される。また個々の患者において疾患活動度と関連する特異的なTCRレパトアを決定することで病勢の予測や治療介入時の指標に利用でき、テーラーメード医療への応用が可能となると考える。

Outline of Final Research Achievements

HTLV-1-infected cells in peripheral blood and CSF cells from HAM patients were targeted for TCR repertoire analysis using molecular barcodes. This analysis revealed a higher clonal expansion in HTLV-1-infected cells and a higher number of HTLV-1 infected cells with polyclonal expansion in HAM patients. The TCR repertoire of infected cells was unique to each patient, and the repertoire signatures were retained over time. Infected cell clones proliferating in the central nervous system showed a different distribution than those in the peripheral blood and were considered to be pathogenic clones that cause inflammation in the CNS of HAM patients.
Our results indicate that TCR repertoire analysis in HTLV-1-infected cells of HAM patients could be used to identify pathogenic clones. Tracking the pathogenic clones longitudinally and across organs is expected as biomarkers for disease monitoring and therapeutic indicators.

Academic Significance and Societal Importance of the Research Achievements

分子バーコードを用いたバイアスのないTCRレパトア解析のパイプラインを確立した。TCRレパトア解析により、HAM患者で病原性のあるHTLV-1感染細胞クローンは個々の患者に特有のものであり、長期に保存されていること、また中枢神経系では末梢血と異なる分布を示すことが判明した。これらの病原性のあるHTLV-1感染細胞クローンの同定・追跡は、疾患活動性の指標や治療介入時のバイオマーカーとして利用できるなど、テーラーメード医療への応用が可能となると考える。

Research Products

• [Journal Article] Immunopathogenic CSF TCR repertoire signatures in virus-associated neurologic disease (2021)
  • Author(s): Nozuma, S. Enose-Akahata, Y. Johnson, K. R. Monaco, M. C. Ngouth, N. Elkahloun, A. Ohayon, J. Zhu, J. Jacobson, S.
  • Journal Title: JCI Insight Volume: 6
  • Peer Reviewed / Open Access
• [Journal Article] Clinical trial of raltegravir, an integrase inhibitor, in HAM/TSP (2021)
  • Author(s): Enose-Akahata, Y. Billioux, B. J. Azodi, S. Dwyer, J. Vellucci, A. Ngouth, N. Nozuma, S. Massoud, R. Cortese, I. Ohayon, J. Jacobson, S.
  • Journal Title: Annals of Clinical and Translational Neurology Volume: 8 Issue: 10 Pages: 1970-1985
  • DOI: 10.1002/acn3.51437
  • Peer Reviewed / Open Access
• [Journal Article] Anti-human T-cell leukemia virus type 1 (HTLV-1) antibody assays in cerebrospinal fluid for the diagnosis of HTLV-1-associated myelopathy/tropical spastic paraparesis (2021)
  • Author(s): Kodama Daisuke, Tanaka Masakazu, Matsuzaki Toshio, Nozuma Satoshi, Matsuura Eiji, Takashima Hiroshi, Izumo Shuji, Kubota Ryuji
  • Journal Title: Journal of Clinical Microbiology Volume: - Issue: 5
  • DOI: 10.1128/jcm.03230-20
  • Peer Reviewed / Open Access
• [Journal Article] Genome wide association study of HTLV-1 associated myelopathy/tropical spastic paraparesis in the Japanese population (2021)
  • Author(s): Penova Marina et al.
  • Journal Title: Proceedings of the National Academy of Sciences Volume: 118 Issue: 11
  • DOI: 10.1073/pnas.2004199118
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] Immunopathogenic CSF TCR repertoire signatures in HAM/TSP (2022)
  • Author(s): Satoshi Nozuma, Yoshimi Enose-Akahata, Kory R. Johnson, Maria Chiara Monaco, Nyater Ngouth, Abdel Elkahloun, Joan Ohayon, Jun Zhu and Steven Jacobson
  • Organizer: 20th International Conference on Human Retrovirology: HTLV and Related Viruses
  • Int'l Joint Research
• [Presentation] 感染細胞を標的としたTCRレパトア解析によるHAMでのバイオマーカー探索 (2022)
  • Author(s): 野妻智嗣、松浦英治、田中正和、児玉大介、松崎敏男、久保田龍二、髙嶋博
  • Organizer: 第63回日本神経学会学術大会
  • Int'l Joint Research
• [Presentation] Immunopathogenic TCR repertoire signature in CSF of virus-associated neurologic disease. (2021)
  • Author(s): Satoshi Nozuma, Yoshimi Enose-Akahata, Kory R. Johnson, Maria Chiara Monaco, Nyater Ngouth, Abdel Elkahloun, Joan Ohayon, Jun Zhu and Steven Jacobson
  • Organizer: 17th International Symposium on NeuroVirology
  • Int'l Joint Research
• [Presentation] Immunopathogenic CSF TCR repertoire signatures in virus-associated neurologic disease (2021)
  • Author(s): Satoshi Nozuma, Eiji Matsuura, Masakazu Tanaka, Toshio Matsuzaki, Ryuji Kubota, Steven Jacobson, Hiroshi Takashima
  • Organizer: 第62回日本神経学会学術大会
  • Int'l Joint Research
• [Presentation] HTLV-1感染細胞を標的としたTCRレパトア解析によるHAMの診断および病勢指標となるバイオマーカーの探索 (2021)
  • Author(s): 野妻智嗣、松浦英治、田中正和、松崎敏男、久保田龍二、髙嶋博
  • Organizer: 第33回日本神経免疫学会学術集会
  • Int'l Joint Research
• [Presentation] HTLV-1感染細胞を標的としたTCRレパトア解析によるHAMの病勢指標となるバイオマーカーの探索 (2021)
  • Author(s): 野妻智嗣、松浦英治、田中正和、松崎敏男、久保田龍二、髙嶋博
  • Organizer: 第7回日本HTLV-1学会学術集会
  • Int'l Joint Research