データベース化を目指したヒトメラノーマMitf-M特異的分子標的療法の開発

• Project/Area Number: 26461698
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Dermatology
• Research Institution: Kyoto Prefectural University of Medicine
• Principal Investigator: 中井 章淳 京都府立医科大学, 医学(系)研究科(研究院), 講師 (80453108)
• Project Period (FY): 2014-04-01 – 2016-03-31
• Project Status: Discontinued (Fiscal Year 2015)
• Budget Amount: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000); Fiscal Year 2016: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2015: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000); Fiscal Year 2014: ¥2,860,000 (Direct Cost: ¥2,200,000、Indirect Cost: ¥660,000)
• Keywords: メラノーマ / 分子治療

Outline of Annual Research Achievements

平成26年度では、CRISPR/Cas9システムを用いてMitfをノックアウトしたところ、ヒトメラノーマの細胞株によっては抗腫瘍効果の得られる可能性が示唆された。平成27年度では、CRISPR/Cas9-Mitfの導入効率と遺伝子発現抑制効果をさらに高めるため、レンチウイルスによりCas9およびguideRNAを発現させるCRISPR/Cas9システムを用いて、ヒトMitfのvariant4を標的としたplasmid(p)Lenti CRISPR/Cas9-Mitfを合成した。In vitroにおいて、pLenti CRISPR/Cas9-Mitfを電気穿孔法でヒトメラノーマ細胞株HMV-IIに導入したところ、コントロール群の8.2%までmRNAレベルでのMitfの発現が抑制された。pLenti CRISPR/Cas9-Mitfを導入したHMV-II群とコントロールHMV-II群において、細胞数と細胞ヴァイアビリティを観察したところ、96時間後では共に有意差(P<0.05）が得られ、pLenti CRISPR/Cas9-Mitfによる細胞増殖抑制効果が確認された。次に、pLenti CRISPR/Cas9-Mitfを導入したHMV-II群とコントロールHMV-II群からRNAを調整し、mRNAレベルでの遺伝子発現を比較検討したところ、Mitf、RIGI、IFN-beta、SOX2、SOX9、STAT1、STAT2、CXCR、c-Myc、CCR1、Hsp70、MDA5、GATA4、Rac1、TFAP2A、TP53、MTUS1、ID2、RUNX1において有意差が得られた。次いで、pLenti CRISPR/Cas9-Mitfを導入したHMV-IIとコントロールHMV-IIを免疫不全マウスに皮下移植して腫瘍の増殖程度を比較したが、両群に有意差はみられなかった。これらの結果から、ヒトメラノーマHMV-IIにおいてMitf遺伝子をノックアウトすると、in vitroでは癌関連遺伝子や種々の遺伝子の発現が抑制され、その結果として細胞増殖抑制効果が得られたと推察された。しかしin vivoでは腫瘍増殖抑制効果はみられなかった。その原因としては、腫瘍を取り巻く環境（血管、神経、膠原線維や弾性線維など）が、腫瘍の増殖を促進させる何らかの影響を与えている可能性があると推察された。

Research Products

• [Journal Article] Reticulate acropigmentation of Kitamura with a novel ADAM10 mutation: a case report. (2016)
  • Author(s): Tsutsumi M, Kono M, Akiyama M, Katoh N, Nakai N.
  • Journal Title: J Dermatol Volume: - Issue: 8 Pages: 1-2
  • DOI: 10.1111/1346-8138.13308
  • Peer Reviewed / Open Access
• [Journal Article] Histology Resembling Cutaneous T-Cell Lymphoma in Nodular Scabies: A Case Report. (2016)
  • Author(s): Kataoka Y, Nakai N, Katoh N.
  • Journal Title: Indian J Dermatol. Volume: 61 Issue: 1 Pages: 125-5
  • DOI: 10.4103/0019-5154.174188
  • Peer Reviewed / Open Access
• [Journal Article] MAGIC Syndrome (Mouth and Genital Ulcers with Inflamed Cartilage): Case Report and Review of the Published Work. (2016)
  • Author(s): Kaneko Y, Nakai N, Kida T, Kawahito Y, Katoh N.
  • Journal Title: Indian J Dermatol. Volume: in press Pages: 1-3
  • Peer Reviewed
• [Journal Article] Multiple Fixed Drug Eruption Caused by Promethazine Methylenedisalicylate as One of the Components of PL Combination Granules (2016)
  • Author(s): Hotta E, Nakai N, Katoh N.
  • Journal Title: Indian J Dermatol. Volume: in press Pages: 1-2
  • Peer Reviewed
• [Journal Article] Acute generalized exanthematous pustulosis caused by dihydrocodeine phosphate in a psoriasis vulgaris patient with a heterozygous IL36RN mutation (2015)
  • Author(s): Nakai N, Sugiura K, Akiyama M, Katoh N
  • Journal Title: JAMA Dermatol Volume: 151 Issue: 3 Pages: 311-315
  • DOI: 10.1001/jamadermatol.2014.3002
  • Peer Reviewed / Open Access
• [Journal Article] Clinical usefulness of Mohs' chemosurgery for palliative purposes in patients with cutaneous squamous cell carcinoma with risk factors or without indication for surgery: three case reports. (2015)
  • Author(s): Nakai N, Okuzawa Y, Katoh N.
  • Journal Title: J Dermatol. Volume: 42 Issue: 4 Pages: 405-7
  • DOI: 10.1111/1346-8138.12767
  • Peer Reviewed / Open Access
• [Journal Article] Melanoma arising from the epidermis overlying an acquired intradermal nevus on the forehead. (2015)
  • Author(s): Nakai N, Katoh N.
  • Journal Title: Indian J Dermatol Venereol Leprol. Volume: 81 Issue: 5 Pages: 533-5
  • DOI: 10.4103/0378-6323.158653
  • Peer Reviewed / Open Access
• [Journal Article] A Case of Inflammatory Linear Verrucous Epidermal Nevus on the Upper Eyelid. (2015)
  • Author(s): Nakai N, Ohshita A, Katoh N.
  • Journal Title: Indian J Dermatol. Volume: 60 Issue: 3 Pages: 323-3
  • DOI: 10.4103/0019-5154.156462
  • Peer Reviewed / Open Access
• [Journal Article] A Case of Gloves and Socks Syndrome and Related Eruptions Caused by Coxsackievirus A4 Infection Mimicking Adult-onset Still's Disease. (2015)
  • Author(s): Ohshita A, Nakai N, Katoh N, Konishi K.
  • Journal Title: Indian J Dermatol. Volume: 60 Issue: 3 Pages: 324-4
  • DOI: 10.4103/0019-5154.156472
  • Peer Reviewed / Open Access
• [Journal Article] Persistent pruritic eruptions in adult-onset Still's disease: additional histopathological findings. (2015)
  • Author(s): Hotta E, Nakai N, Katoh N.
  • Journal Title: Acta Dermatovenerol Croat. Volume: 23 Pages: 146-7
  • Peer Reviewed / Open Access
• [Journal Article] A case of fixed drug eruption caused by loxoprofen sodium hydrate (2015)
  • Author(s): Nakai N, Katoh N.
  • Journal Title: Allergology International Volume: 64 Issue: 4 Pages: 377-378
  • DOI: 10.1016/j.alit.2015.04.007
  • NAID: 130005290279
  • ISSN: 1323-8930, 1440-1592
  • Peer Reviewed / Open Access
• [Journal Article] Severe adult atopic dermatitis successfully treated with concurrent Unseiin and standard treatment (2015)
  • Author(s): Nakai N, Katoh N.
  • Journal Title: Traditional & Kampo Medicine Volume: 2 Issue: 1 Pages: 23-6
  • DOI: 10.1002/tkm2.1011
  • Peer Reviewed / Open Access
• [Journal Article] Anti-tumor effect of Japanese herbal medicine Ninjinto in mouse melanoma: Efficacy of preventive treatment (2015)
  • Author(s): Nakai N, Kishida T, Katoh N
  • Journal Title: Traditional & Kampo Medicine Volume: 2 Issue: 1 Pages: 14-22
  • DOI: 10.1002/tkm2.1010
  • Peer Reviewed / Open Access
• [Journal Article] Adult T-cell Lymphoma Complicated with Epidermodysplasia Verruciformis-like Eruptions. (2015)
  • Author(s): Nakai N, Ohshita A, Kuroda J, Katoh N.
  • Journal Title: Acta Dermatovenerol Croat. Volume: 23 Pages: 304-7
  • Peer Reviewed / Open Access
• [Journal Article] Atypical case of juvenile spring eruption with photosensitivity to a single dose of ultraviolet A irradiation. (2014)
  • Author(s): Ohshita A, Nakai N, Katoh N, Konishi K.
  • Journal Title: J Dermatol. Volume: 41 Issue: 4 Pages: 356-7
  • DOI: 10.1111/1346-8138.12421
  • Peer Reviewed / Open Access
• [Journal Article] A case of cutaneous squamous cell carcinoma arising in an actinic keratosis on the face in a centenarian woman successfully treated with surgery. (2014)
  • Author(s): Yamazato S, Nakai N, Katoh N.
  • Journal Title: Indian J Dermatol. Volume: 59 Issue: 3 Pages: 304-6
  • DOI: 10.4103/0019-5154.131425
  • Peer Reviewed / Open Access
• [Journal Article] Nodular primary localized cutaneous amyloidosis in a patient with pulmonary sarcoidosis. (2014)
  • Author(s): Nakai N, Ozawa A, Katoh N.
  • Journal Title: Indian J Dermatol. Volume: 59 Issue: 3 Pages: 307-8
  • DOI: 10.4103/0019-5154.131430
  • Peer Reviewed / Open Access
• [Journal Article] Maculopapular-Type Drug Eruption Caused by Sitagliptin Phosphate Hydrate: A Case Report and Mini-Review of the Published Work (2014)
  • Author(s): Nakai N, Katoh N.
  • Journal Title: Allergology International Volume: 63 Issue: 3 Pages: 489-491
  • DOI: 10.2332/allergolint.13-LE-0669
  • NAID: 130004687701
  • ISSN: 1323-8930, 1440-1592
  • Peer Reviewed / Open Access
• [Journal Article] Paraneoplastic dermatomyositis associated with gallbladder carcinoma: a case report and mini-review of the published work. (2014)
  • Author(s): Sawada T, Nakai N, Masuda K, Katoh N.
  • Journal Title: Indian J Dermatol. Volume: 59 Issue: 6 Pages: 615-6
  • DOI: 10.4103/0019-5154.143543
  • Peer Reviewed / Open Access
• [Journal Article] Multiple fungal infection in a patient on chronic low dose corticotherapy. (2014)
  • Author(s): Yamazato S, Nakai N, Katoh N.
  • Journal Title: Indian J Dermatol. Volume: 59 Issue: 6 Pages: 636-636
  • DOI: 10.4103/0019-5154.143605
  • Peer Reviewed / Open Access
• [Presentation] フルニエ壊疽の2例 (2015)
  • Author(s): 中川弘己、戸瀬彩加、横瀬千美、若林祐輔、金久史尚、浅井 純、中井章淳、竹中秀也、加藤則人
  • Organizer: 日本皮膚科学会第442回京滋地方会
  • Place of Presentation: 大津市
  • Year and Date: 2015-09-19
• [Presentation] ブロモワレリル尿素によるアナフィラキシーの１例 (2015)
  • Author(s): 田浦麻衣子、山里志穂、金久史尚、田嶋佐妃、中井章淳、浅井 純、益田浩司、加藤則人
  • Organizer: 日本皮膚科学会第442回京滋地方会
  • Place of Presentation: 大津市
  • Year and Date: 2015-09-19
• [Presentation] 網状肢端色素沈着症の１例 (2015)
  • Author(s): 堤 美穂、中井章淳、加藤則人、河野通浩
  • Organizer: 日本皮膚科学会第440回京滋地方会
  • Place of Presentation: 京都市
  • Year and Date: 2015-06-06
• [Presentation] 両手に多発したReticulohistiocytic Granulomaの1例. (2015)
  • Author(s): 上田有希子、中井章淳、加藤則人.
  • Organizer: 第78回日本皮膚科学会東京支部学術大会.
  • Place of Presentation: 東京
  • Year and Date: 2015-02-21 – 2015-02-22
• [Presentation] ワンピースに使用された染料による接触皮膚炎の1例. (2014)
  • Author(s): 田恵理、峠岡理沙、中井章淳、益田浩司、佐々木和実、加藤則人.
  • Organizer: 第44回日本皮膚アレルギー・接触皮膚炎学会総会学術大会.
  • Place of Presentation: 仙台
  • Year and Date: 2014-11-21 – 2014-11-23
• [Book] エキスパートが答える！アトピー性皮膚炎Q&A 55．加藤則人（編） (2014)
  • Author(s): 中井章淳(分担執筆）
  • Total Pages: 2
  • Publisher: 診断と治療社