Establishment of minimally invasive diagnostic method for malignant hyperthermia using fibroblasts

• Project/Area Number: 19K18298
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 55050:Anesthesiology-related
• Research Institution: Hiroshima University
• Principal Investigator: Kondo Takashi 広島大学, 病院(医), 講師 (20711774)
• Project Period (FY): 2019-04-01 – 2022-03-31
• Project Status: Completed (Fiscal Year 2021)
• Budget Amount: ¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000); Fiscal Year 2021: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2020: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2019: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
• Keywords: 悪性高熱症

Outline of Research at the Start

悪性高熱症が疑われる患者に対して診断のための筋生検を行う際に、同時に皮膚から線維芽細胞を採取して培養し、筋芽細胞に分化誘導する遺伝子導入試薬を付加して骨格筋細胞に分化させて機能検査を行う。この診断法が確立すれば従来の検査よりも患者に与える侵襲を軽減することができ、検査が従来より普及することが期待される。また、患者血液の遺伝子情報を解析することで新たな原因遺伝子の同定や病態のさらなる解明も期待される。

Outline of Final Research Achievements

To perform a less invasive functional test for malignant hyperthermia (MH), which requires diagnosis by muscle biopsy, we use a transcription factor (MyoD1) that induces the differentiation of fibroblasts into skeletal muscle cells (myotube cells). We created myotube cells by introducing MyoD1 into skin fibroblasts. Next, a calcium sensitivity test was performed on myotube cells prepared from fibroblasts, and a comparison was made with a calcium susceptibility test on myotube cells in MH patients.
As a result of the examination, the calcium sensitivity of myotube cells derived from MH patients was significantly higher than that of myotube cells prepared from normal fibroblasts. The results of this study showed the possibility that myotube cells prepared from fibroblasts could be used for MH function testing.

Academic Significance and Societal Importance of the Research Achievements

悪性高熱症（MH）の機能診断のために必要な筋生検は侵襲が大きく、特に小児においては全身麻酔が必要となるため検査を行うことが困難な実態にある。
本研究成果により、正常ヒト線維芽細胞を分化させて作成した筋細胞によって機能解析を行うことができることが示された。現状では、MH患者の皮膚線維芽細胞から分化させた筋細胞を用いた機能解析は実用化されていないが、本研究の方法による皮膚を利用する検査法が確立すれば、従来の筋生検を必要とする検査よりも患者に与える侵襲を軽減することができ、MHの検査が従来よりも普及することが期待される。

Research Products

• [Journal Article] Malignant hyperthermia in a 16-day-old infant with congenital diaphragmatic hernia: a case report (2021)
  • Author(s): Tsutsumi YM, Kakuta N, Kawanishi R, Tanaka K, Kanzaki R, Morio A, Noda Y, Miyoshi H, Kondo T, Mukaida K.
  • Journal Title: Journal of anesthesia Volume: 35 Issue: 2 Pages: 311-314
  • DOI: 10.1007/s00540-021-02902-2
  • NAID: 40022524264
  • Peer Reviewed
• [Journal Article] Prediction of blood pressure change during surgical incision under opioid analgesia using sympathetic response evoking threshold (2021)
  • Author(s): Kamiya Satoshi、Nakamura Ryuji、Saeki Noboru、Kondo Takashi、Miyoshi Hirotsugu、Narasaki Soushi、Morio Atsushi、Kawamoto Masashi、Hirano Harutoyo、Tsuji Toshio、Tsutsumi Yasuo M.
  • Journal Title: Scientific Reports Volume: 11 Issue: 1 Pages: 9558-9558
  • DOI: 10.1038/s41598-021-87636-7
  • Peer Reviewed / Open Access
• [Journal Article] Leucine induces cardioprotection in vitro by promoting mitochondrial function via mTOR and Opa-1 signaling. (2021)
  • Author(s): Morio A, Tsutsumi R, Kondo T, Miyoshi H, Kato T, Narasaki S, Satomi S, Nakaya E, Kuroda M, Sakaue H, Kitamura T, Tsutsumi YM
  • Journal Title: Nutr Metab Cardiovasc Dis Volume: 31 Issue: 10 Pages: 2979-2986
  • DOI: 10.1016/j.numecd.2021.06.025
  • Peer Reviewed
• [Journal Article] Leucine imparts cardioprotective effects by enhancing mTOR activity and mitochondrial fusion in a myocardial ischemia/reperfusion injury murine model. (2021)
  • Author(s): Morio A, Tsutsumi R, Satomi S, Kondo T, Miyoshi H, Kato T, Kuroda M, Kitamura T, Hara K, Saeki N, Sakaue H, and Tsutsumi YM
  • Journal Title: Diabetol Metab Syndr Volume: 13 Issue: 1 Pages: 39-39
  • DOI: 10.1186/s13098-021-00755-z
  • Peer Reviewed
• [Journal Article] Intraoperative responses of motor evoked potentials to the novel intravenous anesthetic remimazolam during spine surgery: a report of two cases (2020)
  • Author(s): Kondo Takashi、Toyota Yukari、Narasaki Soshi、Watanabe Tomoyuki、Miyoshi Hirotsugu、Saeki Noboru、Tsutsumi Yasuo M.
  • Journal Title: JA Clinical Reports Volume: 6 Issue: 1 Pages: 97-97
  • DOI: 10.1186/s40981-020-00401-z
  • NAID: 120007169773
  • Peer Reviewed / Open Access
• [Journal Article] Evaluation of a new face mask concept for oxygen administration: a crossover study in healthy volunteers. (2020)
  • Author(s): Kondo T, Saeki N, Otsuki S, Tsutsumi YM.
  • Journal Title: Journal of Anesthesia Volume: 34 Issue: 6 Pages: 950-952
  • DOI: 10.1007/s00540-020-02861-0
  • Peer Reviewed
• [Journal Article] Comparison of the Effects of Desflurane, Sevoflurane, and Propofol on the Glottic Opening Area during Remifentanil-Based General Anesthesia Using a Supraglottic Airway Device (2020)
  • Author(s): Kondo Takashi、Izumi Hiromichi、Kitagawa Makiko
  • Journal Title: Anesthesiology Research and Practice Volume: 2020 Pages: 1-6
  • DOI: 10.1155/2020/1302898
  • Peer Reviewed / Open Access
• [Journal Article] A case of delayed emergence after aortic root and ascending aorta replacement, which was probably caused by portosystemic shunt (2019)
  • Author(s): 呉 晟名, 髙橋 信也, 森田 翔平, 前田 和樹, 片山 桂次郎, 黒崎 達也, 近藤 隆志, 村下 純二, 末田 泰二郎
  • Journal Title: Cardiovascular Anesthesia Volume: 23 Issue: 1 Pages: 83-87
  • DOI: 10.11478/jscva.2019-3-001
  • NAID: 130007709369
  • ISSN: 1342-9132, 1884-7439
  • Year and Date: 2019-08-01
• [Presentation] フォンタン術後患者の副腎外褐色細胞腫に対する後腹膜鏡下腫瘍摘除術の麻酔経験 (2020)
  • Author(s): 近藤隆志，竹野典子，神谷諭史，森尾篤，三好寛二，堤保夫
  • Organizer: 日本心臓血管麻酔学会第25回学術大会
• [Presentation] 悪性高熱症患者で発見された1型リアノジン受容体の一塩基多型E3756Qの機能解析 (2019)
  • Author(s): 近藤隆志，安田季道，神﨑理英子，向田圭子，濱田宏，河本昌志
  • Organizer: 日本麻酔科学会第66回学術集会
• [Presentation] 高度肺高血圧症を伴う心室中隔欠損症および大動脈弁狭窄症に対する根治手術の術後管理に高流量鼻カニュラによる一酸化窒素吸入療法が有用であった1例 (2019)
  • Author(s): 近藤 隆志, 三好 寛二, 濱田宏
  • Organizer: 日本心臓血管麻酔学会第24回学術大会