Annual Report 2017
Division of Molecular and Cellular Medicine
Takahiro Ochiya, Yusuke Yamamoto, Takeshi Katsuda, Yusuke Yoshioka, Ayako Inoue, Kana Kurosaki, Mayuko Yamamura, Nobuyoshi Kosaka, Luc Gailhouste, Ai Hironaka, Takumi Sonoda, Hiroko Tadokoro, Juntaro Matsuzaki, Marta Prieto Vila, Anna Sanchez Calle, Akiko Kogure, Yutaka Naito, Tomomi Imamura, Naomi Nomura, Teruko Yamaguchi, Kazumi Nagao, Megumi Miyagi, Hikaru Sonoda, Satoko Takizawa, Maki Abe, Kurataka Otsuka, Tsukasa Kadota, Junpei Kawauchi, Fumihiko Urabe, Kazunori Hosaka, Masaharu Somiya, Hayato Kurata, Yumi Kawamura, Wataru Usuba, Iwao Shimomura, Isaku Kohama, Tomofumi Yamamoto, Tomoko Yamaguchi, Ryou-u Takahashi, Minami Kumazaki, Makiko Ichikawa, Lee Chuen Liew, Yueyuan Zhou
Introduction
The focus of the Division of Molecular and Cellular Medicine lies in the development of novel treatment and diagnosis for cancer therapy. The specific activities were as follows:
1) An exosome as a novel diagnosis and therapeutic tool against cancer;
2) Drug discovery for targeting miRNA to regulate cancer stem cell property;
3) Development for carcinogenic model using tissue stem cell culture;
4) Small molecule-based reprogramming of adult hepatocytes modelling for hepatocellular carcinomas.
Research activities
1) An exosome as a novel diagnosis and therapeutic tool against cancer
We have identified "Malignant Exosomes" from highly-metastatic ovarian cancer cell line, which remarkably induce the peritoneal dissemination of ovarian cancer cells. Peritoneal mesothelial cells were a barrier of peritoneal cavity and were killed by the malignant exosome due to induced apoptotic cell death. We also found that abundant MMP1 mRNA was packaged in the exosomes and it could be a prognostic marker for early stage ovarian cancer (Yokoi, Nature Commun.). In ovarian cancer, we established serum miRNA biomarkers for the early detection of ovarian cancer. We also reported that most of these miRNAs were packaged in the exosomes (Yokoi, Oncotarget).
Since the exosomes are closely related to cancer metastasis, we have tried to block circulating exosomes in blood using specific antibodies in metastatic breast cancer mouse model (xenograft model). Administration of human specific antibodies into a mouse model significantly reduced metastatic cancer cell numbers in the lung (Nishida-Aoki, Mol. Therapy). Thus, our data clearly demonstrated that targeting the exosomes in the blood of cancer patients is a potential therapeutic strategy for cancer treatment. In addition, we developed a purification protocol for exosomes derived from bovine milk toward drug delivery systems (Somiya, JEV).
In addition to these findings, we have established a rat model to trace exosome secretion using CD63-GFP transgenic rats (Yoshimura, Sci.Rep) and also tried to identify genes involved in exosome secretion, in order to develop a therapeutic method for exosome suppression in cancer cells. We have published to review articles to show the biological significance of exosome in cancer (Kawamura, Cancer science; Kadota, Semin. Cell Dev. Biol.; Naito, CMLS; Liew, Int Immunol).
2) Drug discovery for targeting miRNA to regulate cancer stem cell property
While cancer stem cell (CSC) properties such as tumorigenicity and drug resistance are the major focus in current cancer research, the molecular mechanisms for the regulation of CSC properties are not fully understood. MicroRNA (miRNA) is identified as a target involved in the regulation of CSC properties. We previously identified microRNA-27b (miR-27b) as a key regulator for the generation of a side-population in breast cancer cells that showed CSC properties, and also found that the anti-type II diabetes (T2D) drug metformin reduced this side-population via miR- 27b-mediated repression of ENPP1, which is involved in T2D development. We have begun the development of a mimic (mimetic nucleic acid) delivery system for miR-27b. Using this system, we have confirmed that miR-27b mimic was suc-cessfully delivered into the tumor in tumor-bearing mice. Furthermore, we found that CD44s worked as EMT regulation and modulated drug resistance of cisplatin in head and neck cancer cells (Miyazaki, Oncotarget). In order to search a therapeutic target in breast cancer of young patients, we performed comprehensive miRNA microarray between young and old patients with breast cancer, we established a biomarker predicting prognosis of breast cancer in young women and identified miR-1285-5p as a molecular target (Hironaka-Mitsuhashi A).
3) Development for carcinogenic model using tissue stem cell culture
By applying stable culturing system of human epithelial stem cells (Yamamoto, Inflamm. Regen.), we previously identified stem cell population in pre-cancerous lesion such as Barrett's esophagus which is precursor of esophageal adenocarcinomas. On the basis of the primary cell culture system, we are investigating tissue-specific oncogenic activities and also searching small molecules and drugs that can selectively kill or damage cancer cells which harbor specific mutations.
4) Small molecule-based reprogramming of adult hepatocytes modelling for hepatocellular carcinomas
We are interested in modelling hepatocellular carcinomas utilizing chemically-induced liver progenitor cells. By modifying the technique to reprogram cancer and epithelial cells (Kawamata, BBRC), we have developed a strategy that small molecular cocktail enabled mature hepatocytes to reprogram bi-potential liver progenitor cells at high efficiently. Also, we have established an orthotopic transplantation method for reprogrammed liver progenitor cells which showed remarkably high repopulation rates (Katsuda, Cell Stem Cell; Katsuda, Stem Cell Investig.). Combined these techniques, our main focus is on the elucidation of the multi-step carcinogenesis process of normal liver cells to hepatocellular carcinomas.
Education
We have trained eight doctoral students.
Future prospects
1) Exosome Diagnosis and Treatment Strategy: We aim to develop novel diagnostic methods and new therapeutic methods in various cancer types for exosomes. We filed patent applications for the intellectual property.
2) Cancer stem cells and miRNA: We aim to create new drugs targeting miRNAs that control cancer stem cells.
3) We aim to identify candidate factors for treatment and clarify the nature of cancer by reproducing the process of carcinogenesis of cells from normal cancer cells through precancerous lesions.
4) We will continue elucidating properties of new stem cells by studying hepatic stem cells and mesenchymal stem cells for developing a model of hepatocellular carcinomas.
List of papers published in January 2017 - March 2018
Journal
1. Takahashi RU, Prieto-Vila M, Hironaka A, Ochiya T. The role of extracellular vesicle microRNAs in cancer biology. Clinical chemistry and laboratory medicine, 55:648-656, 2017
2. Matsuzaki J, Ochiya T. Circulating microRNAs and extracellular vesicles as potential cancer biomarkers: a systematic review. Int J Clin Oncol, 22:413-420, 2017
3. Kawamura Y, Yamamoto Y, Sato TA, Ochiya T. Extracellular vesicles as trans-genomic agents: Emerging roles in disease and evolution. Cancer Sci, 108:824-830, 2017
4. Yokoi A, Yoshioka Y, Yamamoto Y, Ishikawa M, Ikeda SI, Kato T, Kiyono T, Takeshita F, Kajiyama H, Kikkawa F, Ochiya T. Malignant extracellular vesicles carrying MMP1 mRNA facilitate peritoneal dissemination in ovarian cancer. Nat Commun, 8:14470, 2017
5. Takano Y, Masuda T, Iinuma H, Yamaguchi R, Sato K, Tobo T, Hirata H, Kuroda Y, Nambara S, Hayashi N, Iguchi T, Ito S, Eguchi H, Ochiya T, Yanaga K, Miyano S, Mimori K. Circulating exosomal microRNA-203 is associated with metastasis possibly via inducing tumor-associated macrophages in colorectal cancer. Oncotarget, 8:78598-78613, 2017
6. Kadota T, Yoshioka Y, Fujita Y, Kuwano K, Ochiya T. Extracellular vesicles in lung cancer-From bench to bedside. Semin Cell Dev Biol, 67:39-47, 2017
7. Uotani K, Fujiwara T, Yoshida A, Iwata S, Morita T, Kiyono M, Yokoo S, Kunisada T, Takeda K, Hasei J, Numoto K, Nezu Y, Yonemoto T, Ishii T, Kawai A, Ochiya T, Ozaki T. Circulating MicroRNA-92b-3p as a Novel Biomarker for Monitoring of Synovial Sarcoma. Sci Rep, 7:14634, 2017
8. Kanda Y, Osaki M, Onuma K, Sonoda A, Kobayashi M, Hamada J, Nicolson GL, Ochiya T, Okada F. Amigo2-upregulation in Tumour Cells Facilitates Their Attachment to Liver Endothelial Cells Resulting in Liver Metastases. Sci Rep, 7:43567, 2017
9. Sanada T, Hirata Y, Naito Y, Yamamoto N, Kikkawa Y, Ishida Y, Yamasaki C, Tateno C, Ochiya T, Kohara M. Transmission of HBV DNA Mediated by Ceramide-Triggered Extracellular Vesicles. Cellular and molecular gastroenterology and hepatology, 3:272-283, 2017
10. De S, Kuwahara S, Hosojima M, Ishikawa T, Kaseda R, Sarkar P, Yoshioka Y, Kabasawa H, Iida T, Goto S, Toba K, Higuchi Y, Suzuki Y, Hara M, Kurosawa H, Narita I, Hirayama Y, Ochiya T, Saito A. Exocytosis-Mediated Urinary Full-Length Megalin Excretion Is Linked With the Pathogenesis of Diabetic Nephropathy. Diabetes, 66:1391-1404, 2017
11. Yokoi A, Yoshioka Y, Hirakawa A, Yamamoto Y, Ishikawa M, Ikeda SI, Kato T, Niimi K, Kajiyama H, Kikkawa F, Ochiya T. A combination of circulating miRNAs for the early detection of ovarian cancer. Oncotarget, 8:89811-89823, 2017
12. Mori H, Suzuki H, Matsuzaki J, Masaoka T, Kanai T. Antibiotic resistance and gyrA mutation affect the efficacy of 10-day sitafloxacin-metronidazole-esomeprazole therapy for Helicobacter pylori in penicillin allergic patients. United European Gastroenterol J, 5:796-804, 2017
13. Matsuzaki J, Suzuki H. Circulating microRNAs as potential biomarkers to detect transformation of Barrett's oesophagus to oesophageal adenocarcinoma. BMJ Open Gastroenterol, 4:e000160, 2017
14. Prieto-Vila M, Takahashi RU, Usuba W, Kohama I, Ochiya T. Drug Resistance Driven by Cancer Stem Cells and Their Niche. Int J Mol Sci, 18:2017
15. Kawamata M, Katsuda T, Yamada Y, Ochiya T. In vitro reconstitution of breast cancer heterogeneity with multipotent cancer stem cells using small molecules. Biochem Biophys Res Commun, 482:750-757, 2017
16. Fujiwara T, Uotani K, Yoshida A, Morita T, Nezu Y, Kobayashi E, Uehara T, Omori T, Sugiu K, Komatsubara T, Takeda K, Kunisada T, Kawamura M, Kawai A, Ochiya T, Ozaki T. Clinical significance of circulating miR-25-3p as a novel diagnostic and prognostic biomarker in osteosarcoma. Oncotarget, 8:33375-33392, 2017
17. Urabe F, Kosaka N, Yoshioka Y, Egawa S, Ochiya T. The small vesicular culprits: the investigation of extracellular vesicles as new targets for cancer treatment. Clinical and translational medicine, 6:45, 2017
18. Kocan B, Maziarz A, Tabarkiewicz J, Ochiya T, Banas-Zabczyk A. Trophic Activity and Phenotype of Adipose Tissue-Derived Mesenchymal Stem Cells as a Background of Their Regenerative Potential. Stem cells international, 2017:1653254, 2017
19. Yamamoto Y, Ochiya T. Epithelial stem cell culture: modeling human disease and applications for regenerative medicine. Inflammation and regeneration, 37:3, 2017
20. Yasui T, Yanagida T, Ito S, Konakade Y, Takeshita D, Naganawa T, Nagashima K, Shimada T, Kaji N, Nakamura Y, Thiodorus IA, He Y, Rahong S, Kanai M, Yukawa H, Ochiya T, Kawai T, Baba Y. Unveiling massive numbers of cancer-related urinary-microRNA candidates via nanowires. Science advances, 3:e1701133, 2017
21. Enomoto Y, Takagi R, Naito Y, Kiniwa T, Tanaka Y, Hamada-Tsutsumi S, Kawano M, Matsushita S, Ochiya T, Miyajima A. Identification of the novel 3' UTR sequences of human IL-21 mRNA as potential targets of miRNAs. Sci Rep, 7:7780, 2017
22. Narita M, Shimura E, Nagasawa A, Aiuchi T, Suda Y, Hamada Y, Ikegami D, Iwasawa C, Arakawa K, Igarashi K, Kuzumaki N, Yoshioka Y, Ochiya T, Takeshima H, Ushijima T, Narita M. Chronic treatment of non-small-cell lung cancer cells with gefitinib leads to an epigenetic loss of epithelial properties associated with reductions in microRNA-155 and -200c. PLoS One, 12:e0172115, 2017
23. Katsuda T, Ochiya T. Biological and clinical insights offered by chemically induced liver progenitors (CLiPs). Stem cell investigation, 4:68, 2017
24. Hironaka-Mitsuhashi A, Matsuzaki J, Takahashi RU, Yoshida M, Nezu Y, Yamamoto Y, Shiino S, Kinoshita T, Ushijima T, Hiraoka N, Shimizu C, Tamura K, Ochiya T. A tissue microRNA signature that predicts the prognosis of breast cancer in young women. PLoS One, 12:e0187638, 2017
25. Ichinohe N, Ishii M, Tanimizu N, Kon J, Yoshioka Y, Ochiya T, Mizuguchi T, Hirata K, Mitaka T. Transplantation of Thy1+ cells accelerates liver regeneration by enhancing the growth of small hepatocyte-like progenitor cells via IL17RB signaling. Stem Cells, 35:920-931, 2017
26. Naito Y, Yoshioka Y, Yamamoto Y, Ochiya T. How cancer cells dictate their microenvironment: present roles of extracellular vesicles. Cell Mol Life Sci, 74:697-713, 2017
27. Katsuda T, Kawamata M, Hagiwara K, Takahashi RU, Yamamoto Y, Camargo FD, Ochiya T. Conversion of terminally committed hepatocytes to culturable bipotent progenitor cells with regenerative capacity. Cell Stem Cell, 20:41-55, 2017
28. Nishida-Aoki N, Tominaga N, Takeshita F, Sonoda H, Yoshioka Y, Ochiya T. Disruption of circulating extracellular vesicles as a novel therapeutic strategy against cancer metastasis. Mol Ther, 25:181-191, 2017
29. Liew LC, Katsuda T, Gailhouste L, Nakagama H, Ochiya T. Mesenchymal stem cell-derived extracellular vesicles: a glimmer of hope in treating Alzheimer's disease. Int Immunol, 29:11-19, 2017
30. Fujita Y, Kadota T, Araya J, Ochiya T, Kuwano K. Extracellular Vesicles: New Players in Lung Immunity. Am J Respir Cell Mol Biol, 58:560-565, 2018
31. Otsuka K, Yamamoto Y, Matsuoka R, Ochiya T. Maintaining good miRNAs in the body keeps the doctor away?: Perspectives on the relationship between food-derived natural products and microRNAs in relation to exosomes/extracellular vesicles. Mol Nutr Food Res, 62:2018
32. Shimagaki T, Yoshizumi T, Harimoto N, Yoshio S, Naito Y, Yamamoto Y, Ochiya T, Yoshida Y, Kanto T, Maehara Y. MicroRNA-125b expression and intrahepatic metastasis are predictors for early recurrence after hepatocellular carcinoma resection. Hepatol Res, 48:313-321, 2018
33. Yoshimura A, Adachi N, Matsuno H, Kawamata M, Yoshioka Y, Kikuchi H, Odaka H, Numakawa T, Kunugi H, Ochiya T, Tamai Y. The Sox2 promoter-driven CD63-GFP transgenic rat model allows tracking of neural stem cell-derived extracellular vesicles. Dis Model Mech, 11:2018
34. Shiozawa K, Shuting J, Yoshioka Y, Ochiya T, Kondo T. Extracellular vesicle-encapsulated microRNA-761 enhances pazopanib resistance in synovial sarcoma. Biochem Biophys Res Commun, 495:1322-1327, 2018
35. Hashimoto K, Ochi H, Sunamura S, Kosaka N, Mabuchi Y, Fukuda T, Yao K, Kanda H, Ae K, Okawa A, Akazawa C, Ochiya T, Futakuchi M, Takeda S, Sato S. Cancer-secreted hsa-miR-940 induces an osteoblastic phenotype in the bone metastatic microenvironment via targeting ARHGAP1 and FAM134A. Proc Natl Acad Sci U S A, 115:2204-2209, 2018
36. Somiya M, Yoshioka Y, Ochiya T. Biocompatibility of highly purified bovine milk-derived extracellular vesicles. J Extracell Vesicles, 7:1440132, 2018
37. Miyazaki H, Takahashi RU, Prieto-Vila M, Kawamura Y, Kondo S, Shirota T, Ochiya T. CD44 exerts a functional role during EMT induction in cisplatin-resistant head and neck cancer cells. Oncotarget, 9:10029-10041, 2018
38. Yamamoto T, Kosaka N, Hattori Y, Ochiya T. A Challenge to Aging Society by microRNA in Extracellular Vesicles: microRNA in Extracellular Vesicles as Promising Biomarkers and Novel Therapeutic Targets in Multiple Myeloma. J Clin Med, 7:2018
39. Gailhouste L, Liew LC, Yasukawa K, Hagiwara K, Iwazaki N, Yamada Y, Hatada I, Ochiya T. Epigenetic Reprogramming of Human Hepatoma Cells: A Low-Cost Option for Drug Metabolism Assessment. Cell Mol Gastroenterol Hepatol, 5:454-457.e1, 2018