Development of genetically modified iPS cell derived M1 macrophage therapy against melanoma

2019 Fiscal Year Research-status Report

• Project/Area Number: 18K08301
• Research Institution: Kumamoto University
• Principal Investigator: 福島 聡 熊本大学, 大学院生命科学研究部(医), 准教授 (50398210)
• Co-Investigator(Kenkyū-buntansha): 尹 浩信 熊本大学, 大学院生命科学研究部(医), 教授 (20282634) [Withdrawn]
• Project Period (FY): 2018-04-01 – 2021-03-31
• Keywords: メラノーマ / 免疫療法 / マクロファージ / iPS細胞

Outline of Annual Research Achievements

B6マウスに同一の遺伝的背景を持つマウスメラノーマを播種させ、同系統のiPS細胞由来マクロファージ（iPS-MP）で治
療する実験系を用いて研究を行った。IL-15、OX40L、4-1BBLによる遺伝子改変、あるいはSTINGリガンドの付与による効果増強を検証した。IL-15、OX40、4-1BBL遺伝子改変を行なったiPS-MPは、遺伝子改変をおこなっていないiPS-MPよりも、有意に腫瘍の播種を抑制した。また、STINGリガンドの付与も同様の傾向を示した。
今年度は特にiPS-MP-OX40L（OX40Lを高発現する遺伝子改変を行ったiPS-MP）の効果発現メカニズム、特に抗原特異的なT細胞に対する影響を解析した。モデル抗原として卵白アルブミン（OVA）とOVA発現マウスメラノーマ（MO4）を用いた。OVAペプチドを負荷したiPS-MP-OX40Lの投与によりマウス体内でOVA特異キラーT細胞が脾臓で増加すること、さらには腫瘍内浸潤も増加することを確認した。腫瘍内浸潤リンパ球の中で制御性T細胞は減少させ、エフェクターT細胞やエフェクターメモリーT細胞は増加させることも確認した。また不死化したiPS細胞をヒトへ応用する前臨床研究として、iPS-MPの腫瘍化や自己免疫現象等の副作用発現がないか検証するために、 B6マウスのiPS-MPにOX40Lを遺伝子導入したもの、あるいはMOCK遺伝子だけを導入したものについて長期経過を観察した。興味深いことに、MHCが同じiPS-MPは腫瘍化すると思われたが、遺伝子改変をしたiPS-MPはより免疫原性が高く排除される傾向にあることがわかった。またこのとき、各臓器に自己免疫現象も起きていないことを組織学的に確認した。

Current Status of Research Progress

2: Research has progressed on the whole more than it was originally planned.

Reason

OX40Lを用いた一連の実験結果については、Pigment cell & melanoma research誌にアクセプトされ、in pressの状態である。他の分子についても、実験は順調に進行している。とくに4-1BBLに関しては、論文作成中の段階である。IL-15と、STINGリガンド負荷については、まだ完全に実験は終了はしていないが、重要な実験成果はすでに得ており、あとは、メカニズムを解析する実験を追加する必要がある。以上より、本研究はおおむね順調に進展していると考える。

Strategy for Future Research Activity

4-1BBL、IL-15、STINGリガンドの実験結果について論文化に入る。さらにはiPS-MPプロジェクトを臨床につなげるために、効果の増強という面から免疫チェックポイント阻害薬との併用も検討する。また安全性を確認するために、MHC一部不一致のアロiPS細胞を用いた実験を行なうこと、安全装置としてのHSV-TKシステムなどの自殺遺伝子の導入することも検討していく。

Research Products

• [Journal Article] Induced pluripotent stem cell-derived myeloid cells expressing OX40 ligand amplify antigen-specific T cells in advanced melanoma. (2020)
  • Author(s): Kimura T, Fukushima S, Okada E, Kuriyama H, Kanemaru H, Kadohisa-Tsuruta M, Kubo Y, Nakahara S, Tokuzumi A, Kajihara I, Makino K, Miyashita A, Aoi J, Makino T, Tsukamoto H, Nishimura Y, Inozume Y, Zhang R, Uemura Y, Senju S, Ihn H.
  • Journal Title: Pigment Cell & Melanoma Research. Volume: - Pages: -
  • Peer Reviewed
• [Journal Article] Inhibition of Endoglin Exerts Antitumor Effects through the Regulation of Non-Smad TGF-β Signaling in Angiosarcoma. (2020)
  • Author(s): Sakamoto R, Kajihara I, Miyauchi H, Maeda-Otsuka S, Yamada-Kanazawa S, Sawamura S, Kanemaru H, Makino K, Aoi J, Makino T, Fukushima S, Masuzawa M, Masuzawa M, Amoh Y, Hoshina D, Abe R, Ihn H.
  • Journal Title: J Invest Dermatol. Volume: - Pages: -
  • Peer Reviewed
• [Journal Article] Successful treatment of occult pancreatic melanoma using BRAF/MEK inhibitors. (2020)
  • Author(s): Mizukami Y, Kanemaru H, Nakamura K, Hashigo S, Kajihara I, Miyashita A, Aoi J, Fukushima S, Honda Y, Ihn H.
  • Journal Title: J Dermatol. Volume: 47 Pages: e126-e127
  • DOI: 10.1111/1346-8138.15198.
  • Peer Reviewed
• [Journal Article] MicroRNAs that predict the effectiveness of anti-PD-1 therapies in patients with advanced melanoma. (2020)
  • Author(s): Nakahara S, Fukushima S, Okada E, Morinaga J, Kubo Y, Tokuzumi A, Matsumoto S, Tsuruta-Kadohisa M, Kimura T, Kuriyama H, Miyashita A, Kajihara I, Jinnin M, Ihn H.
  • Journal Title: J Dermatol Sci. Volume: 97 Pages: 77-79
  • DOI: 10.1016/j.jdermsci.2019.11.010.
  • Peer Reviewed
• [Journal Article] Effect of topical immunotherapy with squaric acid dibutylester for alopecia areata in Japanese patients. (2020)
  • Author(s): Sakai K, Fukushima S, Mizuhashi S, Jinnin M, Makino T, Inoue Y, Ihn H.
  • Journal Title: Allergol Int. Volume: 69 Pages: 274-278
  • DOI: 10.1016/j.alit.2019.10.008.
  • Peer Reviewed / Open Access
• [Journal Article] IL-16 expression is increased in the skin and sera of patients with systemic sclerosis. (2020)
  • Author(s): Kawabata K, Makino T, Makino K, Kajihara I, Fukushima S, Ihn H.
  • Journal Title: Rheumatology (Oxford). Volume: 59 Pages: 519-523
  • DOI: 10.1093/rheumatology/kez318.
  • Peer Reviewed
• [Journal Article] Serious disseminated intravascular coagulation associated with combination therapy of nivolumab and ipilimumab in advanced melanoma. (2020)
  • Author(s): Kuriyama H, Fukushima S, Nakahara S, Kanemaru H, Miyashita A, Aoi J, Tomita Y, Kawasaki T, Nosaka K, Ihn H.
  • Journal Title: J Dermatol. Volume: - Pages: -
  • DOI: 10.1111/1346-8138.15336.
  • Peer Reviewed
• [Journal Article] Remarkable antitumor effect of nivolumab in a patient with metastatic renal cell carcinoma previously treated with a peptidebased vaccine. (2020)
  • Author(s): Kurahashi R, Motoshima T, Fukushima Y, Murakami Y, Yatsuda J, Yamaguchi T, Sugiyama Y, Fukushima S, Komohara Y, Suekane S, Tomomi Kamba T.
  • Journal Title: IJU Case Reports Volume: - Pages: -
  • Peer Reviewed
• [Journal Article] Non-small-cell Lung Cancer with Severe Skin Manifestations Related to Radiation Recall Dermatitis after Atezolizumab Treatment: A Case Report. (2020)
  • Author(s): Nakashima K, Saruwatari K, Sato R, Imamura K, Kajihara I, Fukushima S, Saito T, Ishizuka S, Tamanoi D, Jodai T, Hamada S, Tomita Y, Saeki S, Ichiyasu H, Oya N, Ihn H, Sakagami T.
  • Journal Title: Intern Med. Volume: - Pages: -
• [Journal Article] Onychomycosis caused by Trichosporon cacaoliposimilis. (2020)
  • Author(s): Noguchi H, Matsumoto T, Kimura U, Hiruma M, Kano R, Yaguchi T, Fukushima S, Ihn H.
  • Journal Title: J Dermatol. Volume: - Pages: -
  • DOI: 10.1111/1346-8138.15305.
  • Peer Reviewed
• [Journal Article] Case of metastatic extramammary Paget's disease treated with trastuzumab-biosimilar monotherapy after S-1 and docetaxel combination chemotherapy. (2020)
  • Author(s): Kimura T, Akamatsu Y, Kajihara I, Fukushima S, Ihn H. Ihn H.
  • Journal Title: J Dermatol. Volume: 47 Pages: e1-e2
  • DOI: 10.1111/1346-8138.15096.
  • Peer Reviewed
• [Journal Article] Cytomegalovirus Enterocolitis in a Patient with Refractory Immune-Related Colitis (2020)
  • Author(s): Furuta Y, Miyamoto H, Naoe H, Shimoda M, Hinokuma Y, Miyamura T, Miyashita A, Fukushima S, Tanaka M, Sasaki Y.
  • Journal Title: Case Rep Gastroenterol. Volume: 14 Pages: 103-109
  • DOI: 10.1159/000506186.
  • Peer Reviewed
• [Journal Article] he Warburg effect and tumour immune microenvironment in extramammary Paget's disease: overexpression of lactate dehydrogenase A correlates with immune resistance. (2019)
  • Author(s): Urata K, Kajihara I, Miyauchi H, Mijiddorj T, Otsuka-Maeda S, Sakamoto R, Sawamura S, Kanemaru H, Kanazawa-Yamada S, Makino K, Aoi J, Makino T, Fukushima S, Komohara Y, Ihn H.
  • Journal Title: J Eur Acad Dermatol Venereol. Volume: - Pages: -
  • DOI: 10.1111/jdv.16145.
  • Peer Reviewed
• [Journal Article] Japanese Dermatological Association Guidelines: Outlines of guidelines for cutaneous melanoma 2019. (2019)
  • Author(s): Nakamura Y, Asai J, Igaki H, Inozume T, Namikawa K, Hayashi A, Fukushima S, Fujimura T, Ito T, Imafuku K, Tanaka R, Teramoto Y, Minagawa A, Miyagawa T, Miyashita A, Wada M, Koga H, Sugaya M.
  • Journal Title: J Dermatol. Volume: 47 Pages: 89-103
  • DOI: 10.1111/1346-8138.15151.
  • Peer Reviewed
• [Journal Article] Royal jelly regulates the proliferation of human dermal microvascular endothelial cells through the down-regulation of a photoaging-related microRNA. (2019)
  • Author(s): Kawano Y, Makino K, Jinnin M, Sawamura S, Shimada S, Fukushima S, Ihn H.
  • Journal Title: Drug Discov Ther. Volume: 13 Pages: 268-273
  • DOI: 10.5582/ddt.2019.01070.
  • Peer Reviewed / Open Access
• [Journal Article] Differential predictive factors for cardiovascular events in patients with or without cancer history. (2019)
  • Author(s): Sueta D, Tabata N, Ikeda S, Saito Y, Ozaki K, Sakata K, Matsumura T, Yamamoto-Ibusuki M, Murakami Y, Jodai T, Fukushima S, Yoshida N, Kamba T, Araki E, Iwase H, Fujii K, Ihn H, Kobayashi Y, Minamino T, Yamagishi M, Maemura K, Baba H, Matsui K, Tsujita K.
  • Journal Title: Medicine (Baltimore). Volume: 98 Pages: e17602.
  • DOI: 10.1097/MD.0000000000017602.
  • Peer Reviewed
• [Journal Article] Dermatologic Oncology Group of the Japan Clinical Oncology Group. Confirmatory trial of non-amputative digit preservation surgery for subungual melanoma: Japan Clinical Oncology Group study (JCOG1602, J-NAIL study protocol). (2019)
  • Author(s): Tanaka K, Nakamura Y, Mizutani T, Shibata T, Tsutsumida A, Fukuda H, Matsushita S, Aoki M, Namikawa K, Ohe S, Fukushima S, Yamazaki N
  • Journal Title: BMC Cancer. Volume: 19 Pages: 1002
  • DOI: 10.1186/s12885-019-6248-2.
  • Peer Reviewed / Open Access
• [Journal Article] Type I Interferon Delivery by iPSC-Derived Myeloid Cells Elicits Antitumor Immunity via XCR1+ Dendritic Cells. (2019)
  • Author(s): Tsuchiya N, Zhang R, Iwama T, Ueda N, Liu T, Tatsumi M, Sasaki Y, Shimoda R, Osako Y, Sawada Y, Kubo Y, Miyashita A, Fukushima S, Cheng Z, Nakaki R, Takubo K, Okada S, Kaneko S, Ihn H, Kaisho T, Nishimura Y, Senju S, Endo I, Nakatsura T, Uemura Y.
  • Journal Title: Cell Rep. Volume: 29 Pages: 162-175
  • DOI: 10.1016/j.celrep.2019.08.086.
  • Peer Reviewed / Open Access
• [Journal Article] Japanese real-world study of sequential nivolumab and ipilimumab treatment in melanoma. (2019)
  • Author(s): Tsutsumida A, Fukushima S, Yokota K, Yoshikawa S, Yamasaki O, Tanemura A, Okuyama R, Uhara H, Muto Y, Miyashita A, Akiyama M, Kaji T, Koga H, Kato J, Katayama T, Itakura E, Yamazaki N, Kiyohara Y.
  • Journal Title: J Dermatol. Volume: 46 Pages: 947-955
  • DOI: 10.1111/1346-8138.15073.
  • Peer Reviewed
• [Journal Article] A potential significance of circ_0024169 down regulation in angiosarcoma tissue. (2019)
  • Author(s): Nakashima S, Jinnin M, Ide M, Kajihara I, Igata T, Harada M, Masuguchi S, Fukushima S, Masuzawa M, Masuzawa M, Amoh Y, Ihn H.
  • Journal Title: Intractable Rare Dis Res. Volume: 8 Pages: 129-133
  • DOI: 10.5582/irdr.2019.01034.
  • Peer Reviewed / Open Access
• [Journal Article] Classification of 3097 patients from the Japanese melanoma study database using the American joint committee on cancer eighth edition cancer staging system. (2019)
  • Author(s): Fujisawa Y, Yoshikawa S, Minagawa A, Takenouchi T, Yokota K, Uchi H, Noma N, Nakamura Y, Asai J, Kato J, Fujiwara S, Fukushima S et al.
  • Journal Title: J Dermatol Sci. Volume: 94 Pages: 284-289
  • DOI: 10.1016/j.jdermsci.2019.04.003.
  • Peer Reviewed
• [Journal Article] Clinical and histopathological characteristics and survival analysis of 4594 Japanese patients with melanoma. (2019)
  • Author(s): Fujisawa Y, Yoshikawa S, Minagawa A, Takenouchi T, Yokota K, Uchi H, Noma N, Nakamura Y, Asai J, Kato J, Fujiwara S, Fukushima S et al.
  • Journal Title: Cancer Med. Volume: 8 Pages: 2146-2156
  • DOI: 10.1002/cam4.2110.
  • Peer Reviewed / Open Access
• [Journal Article] Nivolumab-induced colitis in a patient with malignant melanoma: A case report and immunological analysis. (2019)
  • Author(s): Miyashita A, Fukushima S, Tsukamoto H, Itai H, Miyamoto H, Nakahara S, Kubo Y, Kimura T, Kuriyama H, Ihn H.
  • Journal Title: J Dermatol. Volume: 46 Pages: e339-e341
  • DOI: 10.1111/1346-8138.14857.
  • Peer Reviewed
• [Journal Article] Overexpression of cyclin-dependent kinase 4 protein in extramammary Paget's disease. (2019)
  • Author(s): Urata K, Kajihara I, Myangat TM, Tasaki Y, Otsuka-Maeda S, Sawamura S, Kanazawa-Yamada S, Sakamoto R, Makino K, Aoi J, Igata T, Makino T, Masuguchi S, Fukushima S, Jinnin M, Ihn H.
  • Journal Title: J Dermatol. Volume: 46 Pages: 444-448
  • DOI: 10.1111/1346-8138.14858.
  • Peer Reviewed
• [Journal Article] Serum cell-free DNA levels are a useful marker for extramammary Paget disease. (2019)
  • Author(s): Mijiddorj T, Kajihara I, Tasaki Y, Otsuka-Maeda S, Sakamoto R, Sawamura S, Kanazawa-Yamada S, Egashira S, Inoue K, Makino K, Miyashita A, Aoi J, Igata T, Makino T, Masuguchi S, Fukushima S, Jinnin M, Morinaga J, Ikeda T, Ihn H.
  • Journal Title: Br J Dermatol. Volume: 181 Pages: 505-511
  • DOI: 10.1111/bjd.17709.
  • Peer Reviewed
• [Journal Article] Recurrent Fusion Gene ADCK4-NUMBL in Cutaneous Squamous Cell Carcinoma Mediates Cell Proliferation. (2019)
  • Author(s): Egashira S, Jinnin M, Makino K, Ajino M, Shimozono N, Okamoto S, Tazaki Y, Hirano A, Ide M, Kajihara I, Aoi J, Harada M, Igata T, Masuguchi S, Fukushima S, Ihn H.
  • Journal Title: J Invest Dermatol. Volume: 139 Pages: 954-957
  • DOI: 10.1016/j.jid.2018.09.030.
  • Peer Reviewed / Open Access
• [Journal Article] Tinea unguium caused by terbinafine-resistant Trichophyton rubrum successfully treated with fosravuconazole. (2019)
  • Author(s): Noguchi H, Matsumoto T, Hiruma M, Kimura U, Kano R, Yaguchi T, Fukushima S, Ihn H.
  • Journal Title: J Dermatol. Volume: 46 Pages: e446-e447
  • DOI: 0.1111/1346-8138.15033.
  • Peer Reviewed
• [Journal Article] Fungal melanonychia caused by Candida parapsilosis successfully treated with oral fosravuconazole. (2019)
  • Author(s): Noguchi H, Matsumoto T, Kimura U, Hiruma M, Kano R, Yaguchi T, Fukushima S, Ihn H.
  • Journal Title: J Dermatol. Volume: 46 Pages: 911-913
  • DOI: 10.1111/1346-8138.15024.
  • Peer Reviewed
• [Journal Article] Interdigital Hyalohyphomycosis Caused by Members of the Fusarium solani Species Complex. (2019)
  • Author(s): Noguchi H, Matsumoto T, Hiruma M, Kimura U, Yaguchi T, Hirose M, Fukushima S, Ihn H.
  • Journal Title: Acta Derm Venereol. Volume: 99 Pages: 835-836
  • DOI: 10.2340/00015555-3202.
  • Peer Reviewed / Open Access
• [Journal Article] Natural course of epidermolysis bullosa simplex with mottled pigmentation in a Japanese family with the p.P25L mutation in KRT5. (2019)
  • Author(s): Okamura K, Fukushima S, Yamashita J, Abe Y, Hayashi M, Hozumi Y, Ihn H, Suzuki T
  • Journal Title: J Dermatol. Volume: 46 Pages: e233-e235
  • DOI: 10.1111/1346-8138.14788.
  • Peer Reviewed
• [Journal Article] Case of cutaneous myoepithelioma managed with surgical resection without recurrence for 4 years. (2019)
  • Author(s): Kanemaru H, Honda N, Kubo Y, Nishihara H, Onuma T, Sakai K, Fukushima S, Inoue Y, Honda Y, Ihn H.
  • Journal Title: J Dermatol. Volume: 46 Pages: e206-e208
  • DOI: 10.1111/1346-8138.14769.
  • Peer Reviewed
• [Presentation] Combination of anti-IL-6 and anti-PD-L1 antibodies synergistically induced a potent cancer immunity by activating both CTL and Th1 cells in mice. (2019)
  • Author(s): Nishimura Y, Fujieda K, Miyashita A, Fukushima S, Ikeda T, Kubo Y, Senju S, Ihn H, Oshiumi H, Tsukamoto H.
  • Organizer: AACR (American Association for Cancer Research)
  • Int'l Joint Research