遺伝子改変ヒトiPS細胞由来マクロファージを用いた悪性黒色腫に対する免疫療法

2014 Fiscal Year Annual Research Report

• Project/Area Number: 25870545
• Research Institution: Kumamoto University
• Principal Investigator: 福島 聡 熊本大学, 医学部附属病院, 講師 (50398210)
• Project Period (FY): 2013-04-01 – 2015-03-31
• Keywords: 癌 / 免疫学 / 悪性黒色腫 / iPS細胞 / マクロファージ / microRNA / 癌抗原 / 免疫療法

Outline of Annual Research Achievements

研究代表者らのグループは、ヒトiPS細胞から免疫調整機能を持った樹状細胞および抗腫瘍効果を有するマクロファージへの分化誘導に成功していた。強力な抗腫瘍効果を発揮するように遺伝子改変したiPS細胞由来マクロファージ(iPS-MP)を用いる計画であった。これまでに得たデータにより、iPS-MP治療には大量の細胞数(マウス一匹あたり1回の治療で1千万個)を必要とすることがわかっていた。将来、汎用化治療として臨床応用を実現化するためには、安定した治療細胞の準備不可欠である。そこでヒトiPS細胞から分化誘導した骨髄系前駆細胞に、cMYC+BMI1/MDM2を導入して不死化し、iPS細胞由来ミエロイドライン(iPS-ML)を樹立。その後にマクロファージ（iPS-MP）や樹状細胞（iPS-DC）を誘導し、治療に用いる方略をとった。まずiPS-MLの段階で、IFN-αあるいはβといったメラノーマに対する抗腫瘍活性が証明されている遺伝子の導入を行った。それを用いてiPS-MPを作製し、メラノーマに対する抗腫瘍実験を行った。免疫不全マウスにヒトメラノーマ細胞を腹膜播種させた後、種々の遺伝子改変iPS-MPで治療する実験を行なった。結果としてIFN-αあるいはβ遺伝子導入iPS-MPは非常に強力な抗腫瘍効果を示した。さらに抗腫瘍メカニズムを解明を行った。その結果、IFN-αあるいはβの遺伝子導入したiPS-MPは、遺伝子導入していないiPS-MLに比べM1マクロファージと呼ばれる抗腫瘍に機能するタイプに変化することを初めて見出した。マイクロアレイ解析から見出した癌特異抗原（KIF20A, FOXM1）について、ヒトメラノーマ組織での発現を確認し、その機能解析を行なった。また血清マーカーとしてのmicroRNAについても検討し、miR-221が早期診断に有用なこと、6つのmiRの組み合わせ（miR-9, miR-145, miR-150, miR-155, miR-203,miR-205）が転移巣の検出に有用であることを報告した。

Research Products

• [Journal Article] Hair shaft miRNA-221 levels as a new tumor marker of malignant melanoma (2015)
  • Author(s): Inada T, Fukushima S, Murai M, Jinnin M, Miyashita A, Nakahara S, Yamashita J, Aoi J, Masuguchi S, Ihn H.
  • Journal Title: J Dermatol. Volume: 42 Pages: 198-201
  • DOI: 10.1111/1346-8138.12730.
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Combination chemotherapy with S-1 and docetaxel for cutaneous angiosarcoma resistant to paclitaxel. (2015)
  • Author(s): Kajihara I, Kanemaru H, Miyake T, Aoi J, Masuguchi S, Fukushima S, Jinnin M, Ihn H.
  • Journal Title: Drug Discov Ther. Volume: 9 Pages: 75-77
  • DOI: 10.5582/ddt.2015.01005.
  • Peer Reviewed
• [Journal Article] Evaluation of sentinel node biopsy for cutaneous squamous cell carcinoma (2014)
  • Author(s): Fukushima S, Masuguchi S, Igata T, Harada M, Aoi J, Miyashita A, Nakahara S, Inoue Y, Jinnin M, Shiraishi S, Yamashita Y, Ishihara T, Ihn H.
  • Journal Title: J Dermatol. Volume: 41 Pages: 539-541
  • DOI: 10.1111/1346-8138.12508.
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Aesthetic reconstruction of defects in the lateral side wall of the nose using a local V-Y flap. (2014)
  • Author(s): Aoi J, Masuguchi S, Fukushima S, Jinnin M, Ihn H.
  • Journal Title: J Dermatol. Volume: 41 Pages: 1026-1028
  • DOI: 10.1111/1346-8138.12660.
  • Peer Reviewed
• [Journal Article] A simple and effective reconstructive technique for repairing defects of the upper lip using a hatchet flap. (2014)
  • Author(s): Aoi J, Nagamoto E, Masuguchi S, Fukushima S, Jinnin M, Ihn H.
  • Journal Title: J Dermatol. Volume: 41 Pages: 1025-1026
  • DOI: 10.1111/1346-8138.12645.
  • Peer Reviewed
• [Journal Article] A case of anaplastic large cell lymphoma of skeletal muscle. (2014)
  • Author(s): Kubo Y, Aoi J, Johno T, Makino T, Sakai K, Masuguchi S, Fukushima S, Jinnin M, Inoue Y, Ihn H.
  • Journal Title: J Dermatol. Volume: 41 Pages: 999-1002
  • DOI: 10.1111/1346-8138.12641.
  • Peer Reviewed
• [Journal Article] Down-regulation of microRNA-196a in the sera and involved skin of localized scleroderma patients. (2014)
  • Author(s): Makino T, Jinnin M, Etoh M, Yamane K, Kajihara I, Makino K, Ichihara A, Igata T, Sakai K, Fukushima S, Ihn H.
  • Journal Title: Eur J Dermatol. Volume: 24 Pages: 470-476
  • DOI: 10.1684/ejd.2014.2384.
  • Peer Reviewed
• [Journal Article] Clinical significance of serum vascular endothelial-cadherin levels in inflammatory skin diseases. (2014)
  • Author(s): Kuriyama H, Jinnin M, Kanemaru H, Ichihara A, Fujisawa A, Moriya C, Fukushima S, Ihn H.
  • Journal Title: Ann Dermatol. Volume: 26 Pages: 536-538
  • DOI: 10.5021/ad.2014.26.4.536.
  • Peer Reviewed
• [Journal Article] Serum levels of soluble carbonic anhydrase IX are decreased in patients with diffuse cutaneous systemic sclerosis compared to those with limited cutaneous systemic sclerosis. (2014)
  • Author(s): Makino K, Jinnin M, Makino T, Kajihara I, Fukushima S, Inoue Y, Ihn H.
  • Journal Title: Biosci Trends. Volume: 8 Pages: 144-148
  • Peer Reviewed / Open Access
• [Journal Article] The expression profile of the toll-like receptor family in scleroderma dermal fibroblasts. (2014)
  • Author(s): Sakoguchi A, Nakayama W, Jinnin M, Wang Z, Yamane K, Aoi J, Makino K, Kajihara I, Ichihara A, Makino T, Fukushima S, Sakai K, Inoue Y, Ihn H.
  • Journal Title: Clin Exp Rheumatol. Volume: 32 Pages: 4-9
  • Peer Reviewed
• [Journal Article] Platelet-rich plasma therapy is effective for the treatment of refractory skin ulcers in patients with systemic sclerosis. (2014)
  • Author(s): Kanemaru H, Kajihara I, Yamanaka K, Igata T, Makino T, Masuguchi S, Fukushima S, Jinnin M, Ihn H.
  • Journal Title: Mod Rheumatol. Volume: 16 Pages: 1-2
  • Peer Reviewed
• [Journal Article] Immunoglobulin G4-related disease in a psoriasis vulgaris patient treated with ustekinumab. (2014)
  • Author(s): Kajihara I, Makino K, Ichihara A, Fukushima S, Jinnin M, Hirooka S, Kojima K, Kourogi H, Ihn H.
  • Journal Title: J Dermatol. Volume: 41 Pages: 670-671
  • DOI: 10.1111/1346-8138.12528.
  • Peer Reviewed
• [Journal Article] Knockout of endothelial cell-derived endothelin-1 attenuates skin fibrosis but accelerates cutaneous wound healing. (2014)
  • Author(s): Makino K, Jinnin M, Aoi J, Kajihara I, Makino T, Fukushima S, Sakai K, Nakayama K, Emoto N, Yanagisawa M, Ihn H.
  • Journal Title: PLoS One. Volume: 22 Pages: e97972
  • DOI: 10.1371/journal.pone.0097972.
  • Peer Reviewed
• [Journal Article] Increased CCL22 expression in psoriatic skin predicts a good response to infliximab therapy. (2014)
  • Author(s): Kusumoto S, Kajihara I, Nagamoto E, Makino K, Ichihara A, Aoi J, Johno T, Makino T, Fukushima S, Jinnin M, Ihn H.
  • Journal Title: Br J Dermatol. Volume: 175 Pages: 1259-1261
  • DOI: 10.1111/bjd.13091.
  • Peer Reviewed
• [Journal Article] Case of disseminated cutaneous Mycobacterium chelonae infection mimicking cutaneous vasculitis. (2014)
  • Author(s): Ichihara A, Jinnin M, Fukushima S, Inoue Y, Ihn H.
  • Journal Title: J Dermatol. Volume: 41 Pages: 414-417
  • DOI: 10.1111/1346-8138.12459.
  • Peer Reviewed
• [Journal Article] miR-424 levels in hair shaft are increased in psoriatic patients. (2014)
  • Author(s): Tsuru Y, Jinnin M, Ichihara A, Fujisawa A, Moriya C, Sakai K, Fukushima S, Ihn H.
  • Journal Title: J Dermatol. Volume: 41 Pages: 382-385
  • DOI: 10.1111/1346-8138.12460.
  • Peer Reviewed
• [Journal Article] Case of Fusarium paronychia successfully treated with occlusive dressing of antifungal cream. (2014)
  • Author(s): Ikeda I, Ohno T, Ohno H, Miyazaki Y, Nishimoto K, Fukushima S, Makino T, Ihn H.
  • Journal Title: J Dermatol. Volume: 41 Pages: 340-342
  • DOI: 10.1111/1346-8138.12420
  • Peer Reviewed
• [Journal Article] miR-205 down-regulation promotes proliferation of dermatofibrosarcoma protuberans tumor cells by regulating LRP-1 and ERK phosphorylation. (2014)
  • Author(s): Kajihara I, Jinnin M, Harada M, Makino K, Honda N, Makino T, Igata T, Masuguchi S, Fukushima S, Ihn H.
  • Journal Title: Arch Dermatol Res. Volume: 306 Pages: 367-374
  • DOI: 10.1007/s00403-014-1452-z.
  • Peer Reviewed
• [Journal Article] Analysis of expression pattern of serum microRNA levels in patients with psoriasis. (2014)
  • Author(s): Koga Y, Jinnin M, Ichihara A, Fujisawa A, Moriya C, Sakai K, Fukushima S, Inoue Y, Ihn H.
  • Journal Title: J Dermatol Sci. Volume: 74 Pages: 170-171
  • DOI: 10.1016/j.jdermsci.2014.01.005.
  • Peer Reviewed
• [Journal Article] Decreased interleukin-20 expression in scleroderma skin contributes to cutaneous fibrosis. (2014)
  • Author(s): Kudo H, Jinnin M, Asano Y, Trojanowska M, Nakayama W, Inoue K, Honda N, Kajihara I, Makino K, Fukushima S, Ihn H.
  • Journal Title: Arthritis Rheumatol. Volume: 66 Pages: 1636-1647
  • DOI: 10.1002/art.38380.
  • Peer Reviewed
• [Journal Article] The proportion of lymphocytic inflammation with CD123-positive cells in lupus erythematous profundus predict a clinical response to treatment. (2014)
  • Author(s): Miyashita A, Fukushima S, Makino T, Yoshino Y, Yamashita J, Honda N, Aoi J, Ichihara A, Jinnin M, Inoue Y, Ihn H.
  • Journal Title: Acta Derm Venereol. Volume: 94 Pages: 563-567
  • DOI: 10.2340/00015555-1777.
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Upregulation of miR-18a-5p contributes to epidermal necrolysis in severe drug eruptions. (2014)
  • Author(s): Ichihara A, Wang Z, Jinnin M, Izuno Y, Shimozono N, Yamane K, Fujisawa A, Moriya C, Fukushima S, Inoue Y, Ihn H.
  • Journal Title: J Allergy Clin Immunol. Volume: 133 Pages: 1065-1074
  • DOI: 10.1016/j.jaci.2013.09.019.
  • Peer Reviewed
• [Presentation] Serum leptin receptor as a tumor marker of melanoma (2014)
  • Author(s): Satoshi Fukushima, Hironori Mizutani, Azusa Miyashita, Junji Yamashita, Satoshi Nakahara, Masatoshi Jinnin, Hironobu Ihn
  • Organizer: The 39th Annual meeting of the Japanese Society for Investigative Dermatology
  • Place of Presentation: 阪急エキスポパーク、大阪府吹田市
  • Year and Date: 2014-12-12 – 2014-12-14
• [Presentation] 限局性強皮症の血清マーカーとしてのSPARC (2014)
  • Author(s): 福島聡、牧野貴充、神人正寿、尹浩信
  • Organizer: 第42回日本臨床免疫学会総会
  • Place of Presentation: 京王プラザ、東京都新宿区
  • Year and Date: 2014-09-25 – 2014-09-27
• [Presentation] Serum levels of leptin receptor as a new tumor marker of malignant melanoma (2014)
  • Author(s): Satoshi Fukushima, Hironori Mizutani, Azusa Miyashita, Junji Yamashita, Satoshi Nakahara, Masatoshi Jinnin and Hironobu Ihn
  • Organizer: 44th Annual ESDR meeting
  • Place of Presentation: Tivoli Congress Center, Copenhagen, Denmark
  • Year and Date: 2014-09-10 – 2014-09-13
• [Presentation] ついに免疫療法の時代が到来！？イピリムマブの登場 (2014)
  • Author(s): 福島聡
  • Organizer: 第52回日本癌治療学会学術集会
  • Place of Presentation: パシフィコ横浜、横浜市
  • Year and Date: 2014-08-28 – 2014-08-30
  • Invited
• [Presentation] 「メラノーマ：薬物療法」 ついに免疫療法の時代が来る！ (2014)
  • Author(s): 福島聡
  • Organizer: 第30回日本皮膚悪性腫瘍学会学術大会
  • Place of Presentation: 如水会館、東京都千代田区
  • Year and Date: 2014-07-04 – 2014-07-05
  • Invited
• [Presentation] メラノーマに対する遺伝子改変ヒトiPS細胞由来ミエロイドラインによる免疫細胞療法 (2014)
  • Author(s): 福島聡、宮下梓、千住覚、尹浩信
  • Organizer: 第35回日本炎症・再生医学会
  • Place of Presentation: 万国津梁館、沖縄県名護市
  • Year and Date: 2014-07-01 – 2014-07-04
• [Presentation] アトピー性皮膚炎と バリア異常 (2014)
  • Author(s): 福島聡
  • Organizer: 第12回南九州地区合同皮膚科地方会
  • Place of Presentation: 森都心プラザ、熊本市
  • Year and Date: 2014-06-28 – 2014-06-29
  • Invited
• [Presentation] Sentinel node biopsy for non-melanoma skin cancers: A review of 88 cases at one institution (2014)
  • Author(s): Satoshi Fukushima, Hironobu Ihn
  • Organizer: 11th Meeting of the German-Japanese Society of Dermatology
  • Place of Presentation: Marriott Hotel, Heidelberg, Germany
  • Year and Date: 2014-06-11 – 2014-06-14
• [Presentation] Clinical significance of CD169-positive lymph node macrophages in human malignant tumors. (2014)
  • Author(s): Koji Ohnishi, Yoichi Saito, Yoshihiro Komohara, Yuji Miyamoto, Satoshi Fukushima, Munekage Yamaguchi, Hideo Baba, Hironobu Ihn, Hidetaka Katabuchi, Motohiro Takeya
  • Organizer: ASCO 2014
  • Place of Presentation: McCormick Place, Chicago, Illinois
  • Year and Date: 2014-05-30 – 2014-06-03
• [Book] 養子免疫療法、樹状細胞療法、ワクチン療法 皮膚科臨床アセット 17 皮膚の悪性腫瘍 (2014)
  • Author(s): 福島聡
  • Total Pages: 364(78-86)
  • Publisher: 中山書店
• [Book] 「Clinical trials」既存治療無効の進行期メラノーマに対するイピリムマブの第III相試験 Immune-Oncology Frontier (2014)
  • Author(s): 福島聡
  • Total Pages: 60(46-48)
  • Publisher: メディカルレビュー社