Investigation of gene mutation specific to canine bone marrow diseases

• Project/Area Number: 18K05967
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 42020:Veterinary medical science-related
• Research Institution: The University of Tokyo
• Principal Investigator: Goto-Koshino Yuko 東京大学, 大学院農学生命科学研究科(農学部), 特任准教授 (80436518)
• Project Period (FY): 2018-04-01 – 2021-03-31
• Project Status: Completed (Fiscal Year 2020)
• Budget Amount: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2020: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2019: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000); Fiscal Year 2018: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
• Keywords: 真性多血症 / 犬 / 網羅的解析 / クローン性

Outline of Final Research Achievements

Investigation of a polycythemia vera (PV)-specific gene mutation was performed using two dogs diagnosed as PV. CD3 positive lymphocytes were purified as normal cell population, and the remaining CD3 negative nucleated cells were separated as population containing abnormal clone. Two populations were compared to find abnormal clone-specific gene mutation.　Whole exome sequencing was designed at first, however, because of the low quality of genome DNA purified from CD3 positive normal cell population, target resequencing of the cancer related genes was adopted instead. Multiple gene mutations specific to the population containing abnormal clone were extracted, however, none of the mutation was validated by Sanger sequencing. It would be necessary to enrich the abnormal clone, for example, purification of erythroid progenitors from PV dogs, for further analysis.

Academic Significance and Societal Importance of the Research Achievements

真性多血症は血液幹細胞の異常によって異常な赤血球増多が起こる疾患である。異常な血液幹細胞は赤血球だけでなく白血球にも分化することが知られているが、白血病と異なり、細胞の機能は正常である。
本研究は犬における真性多血症の原因遺伝子を探索し、人の真性多血症において高頻度に認められるJAK2遺伝子変異と同様に診断マーカーとして確立すること、さらに治療のターゲットとしての可能性を検討することを目的として行ったが、研究期間内には有望な遺伝子変異を見出すことができなかった。今後は異常細胞のみを単離して解析を試みる予定である。

Research Products

• [Journal Article] Measurement of the concentration of serum soluble interleukin-2 receptor alpha chain in dogs with lymphoma (2020)
  • Author(s): Mizutani Noriyuki、Goto-Koshino Yuko、Kurata Keigo、Fujiwara-Igarashi Aki、Sakaguchi Masahiro、Asada Minoru、Ohno Koichi、Tsujimoto Hajime
  • Journal Title: Veterinary Immunology and Immunopathology Volume: 225 Pages: 110054-110054
  • DOI: 10.1016/j.vetimm.2020.110054
  • Peer Reviewed
• [Journal Article] Comparison of the efficacy of cyclosporine and leflunomide in treating inflammatory colorectal polyps in miniature dachshunds (2020)
  • Author(s): Tani A、Seno T、Yokoyama N、Nakagawa T、Tomiyasu H、Goto-Koshino Y、Tsujimoto H、Ohno K
  • Journal Title: Journal of Veterinary Medical Science Volume: 82 Issue: 4 Pages: 437-440
  • DOI: 10.1292/jvms.19-0560
  • NAID: 130007829876
  • ISSN: 0916-7250, 1347-7439
  • Peer Reviewed / Open Access
• [Journal Article] ErbB2 Copy Number Aberration in Canine Urothelial Carcinoma Detected by a Digital Polymerase Chain Reaction Assay. (2020)
  • Author(s): Sakai K, Maeda S, Saeki K, Yoshitake R, Goto-Koshino Y, Nakagawa T, Nishimura R, Yonezawa T, Matsuki N
  • Journal Title: Vet Pathol Volume: 57 Issue: 1 Pages: 56-65
  • DOI: 10.1177/0300985819879445
  • Peer Reviewed
• [Journal Article] Development of canine X-chromosome inactivation pattern analysis for the detection of cell clonality by incorporating the examination of the SLIT and NTRK-like family member 4 (SLITRK4) gene. (2019)
  • Author(s): Tomita A, Mochizuki H, Tsuboi M, Ogura I, Igarashi H, Goto-Koshino Y, Takahashi M, Ohmi A, Tomiyasu H, Ohno K, Nakagawa T, Uchida K, Nishimura R, Tsujimoto H
  • Journal Title: Res Vet Sci Volume: 125 Pages: 170-175
  • DOI: 10.1016/j.rvsc.2019.06.004
  • Peer Reviewed
• [Journal Article] Variations in ATP7B in cats with primary copper-associated hepatopathy. (2019)
  • Author(s): Asada H, Chambers JK, Kojima M, Goto-Koshino Y, Kanagawa T, Yokoyama N, Tsuboi M, Uchida K, Tsujimoto H, Ohno K
  • Journal Title: J Feline Med Surg Volume: - Issue: 8 Pages: 753-759
  • DOI: 10.1177/1098612x19884763
  • Peer Reviewed
• [Journal Article] Evaluation of visceral fat mass in dogs by computed tomography (2019)
  • Author(s): Nagao I, Ohno K, Nagahara T, Yokoyama N, Nakagawa T, Fujiwara R, Yamamoto K, Goto-Koshino Y, Tomiyasu H, Tsujimoto H
  • Journal Title: Journal of Veterinary Medical Science Volume: 81 Issue: 11 Pages: 1552-1557
  • DOI: 10.1292/jvms.19-0254
  • NAID: 130007744339
  • ISSN: 0916-7250, 1347-7439
  • Peer Reviewed
• [Journal Article] Effect of a two-base insertion mutation of the TP53 gene on expression of p53 protein in canine histiocytic sarcoma cells. (2019)
  • Author(s): Asada H, Toiyasu H, Goto-Koshino Y, Ohno K, Tsujimoto H
  • Journal Title: Am J Vet Res Volume: 80 Issue: 7 Pages: 680-688
  • DOI: 10.2460/ajvr.80.7.680
  • Peer Reviewed
• [Journal Article] Comprehensive analysis of miRNA and protein profiles within exosomes derived from canine lymphoid tumour cell lines. (2019)
  • Author(s): Asada H, Yomiyasu H, Uchikai T, Ishihara G, Goto-Koshino Y, Ohno K, Tsujimoto H
  • Journal Title: PLoS One Volume: 14 Issue: 4 Pages: e0208567-e0208567
  • DOI: 10.1371/journal.pone.0208567
  • Peer Reviewed / Open Access
• [Journal Article] Clinical significance of the two-base insertion mutation in the TP53 gene in canine histiocytic sarcoma. (2019)
  • Author(s): Asada H, Tomiyasu H, Okada K, Chambers JK, Goto-Koshino Y, Uchida K, Kagawa Y, Ohno K, Tsujimoto H.
  • Journal Title: Res Vet Sci Volume: 124 Pages: 57-60
  • DOI: 10.1016/j.rvsc.2019.03.003
  • Peer Reviewed
• [Journal Article] The intratumor heterogeneity of <i>TP53</i> gene mutations in canine histiocytic sarcoma (2019)
  • Author(s): Asada H, Ichii O, Tomiyasu H, Uchida K, Chambers JK, Goto-Koshino Y, Ohno K, Kon Y, Tsujimoto H.
  • Journal Title: Journal of Veterinary Medical Science Volume: 81 Issue: 3 Pages: 353-356
  • DOI: 10.1292/jvms.18-0419
  • NAID: 130007612993
  • ISSN: 0916-7250, 1347-7439
  • Peer Reviewed / Open Access
• [Journal Article] Hepatic copper accumulation in a young cat with familial variations in the ATP7B gene. (2019)
  • Author(s): Asada H, Kojima M, Nagahara T, Goto-Koshino Y, Chambers JK, Nakagawa T, Yokoyama N, Uchida K, Tsujimoto H, Ohno K.
  • Journal Title: J Vet Intern Med Volume: 33 Issue: 2 Pages: 874-878
  • DOI: 10.1111/jvim.15399
  • Peer Reviewed
• [Journal Article] A case of hemophagocytic syndrome progressing into large granular lymphoma in a dog. (2019)
  • Author(s): Akiyoshi M, Hisasue M, Neo S, Akiyoshi M, Goto-Koshino Y.
  • Journal Title: Vet Clin Pathol Volume: 48 Issue: 1 Pages: 71-77
  • DOI: 10.1111/vcp.12704
  • Peer Reviewed
• [Journal Article] Immunohistochemical Profiling of Canine Intestinal T-Cell Lymphomas. (2019)
  • Author(s): Matsumoto I, Nakashima K, Goto-Koshino Y, Chambers JK, Tsujimoto H, Nakayama H, Uchida K.
  • Journal Title: Vet Pathol Volume: 56 Issue: 1 Pages: 50-60
  • DOI: 10.1177/0300985818800015
  • Peer Reviewed
• [Journal Article] Effect of prednisolone administration on gallbladder emptying rate and gallbladder bile composition in dogs. (2018)
  • Author(s): Nagahara T, Ohno K, Kanemoto H, Kakimoto T, Fukushima K, Goto-Koshino Y, Tsujimoto H.
  • Journal Title: Am J Vet Res Volume: 79 Issue: 10 Pages: 1050-1056
  • DOI: 10.2460/ajvr.79.10.1050
  • Peer Reviewed
• [Journal Article] Clinical and clinicopathological characteristics of acute lymphoblastic leukaemia in six cats. (2018)
  • Author(s): Tomiyasu H, Doi A, Chambers JK, Goto-Koshino Y, Ohmi A, Ohno K, Tsujimoto H.
  • Journal Title: J Small Anim Pract Volume: 59 Issue: 12 Pages: 742-746
  • DOI: 10.1111/jsap.12917
  • Peer Reviewed
• [Journal Article] Prognostic value of CD44 variant isoform expression in dogs with multicentric high-grade B-cell lymphoma. (2018)
  • Author(s): Motegi T, Tomiyasu H, Goto-Koshino Y, Takahashi M, Hiyoshi-Kanemoto S, Fujino Y, Ohno K, Tsuimoto H.
  • Journal Title: Am J Vet Res Volume: 79 Issue: 9 Pages: 961-969
  • DOI: 10.2460/ajvr.79.9.961
  • Peer Reviewed
• [Journal Article] Pathological features of intestinal T‐cell lymphoma in Shiba dogs in Japan (2018)
  • Author(s): Matsumoto I., Uchida, K., Nakashima, K., Goto-Koshino, Y., Chambers, J., Tsujimoto, H. and Nakayama, H.
  • Journal Title: Veterinary and Comparative Oncology Volume: 印刷中 Issue: 4 Pages: 417-423
  • DOI: 10.1111/vco.12396
  • Peer Reviewed
• [Journal Article] Prognostic significance of hypermethylation of death-associated protein kinase (DAPK) gene CpG island in dogs with high-grade B-cell lymphoma (2018)
  • Author(s): Sato, M., Mochizuki, H., Goto-Koshino, Y., Fujiwara-Igarashi, A., Takahashi, M., Ohno, K. and Tsujimoto, H.
  • Journal Title: Veterinary and Comparative Oncology Volume: 印刷中 Issue: 3 Pages: 409-415
  • DOI: 10.1111/vco.12395
  • Peer Reviewed