コーンビームCTによる画像誘導放射線治療を併用した前立腺癌に対する強度変調回転照射の検討

学位の種別: 課程博士審査委員会委員 : (主査)東京大学教授 宮川 清, 東京大学准教授 桐生 茂, 東京大学准教授 朝蔭 孝宏, 東京大学准教授 福原 浩, 東京大学講師 山下 英臣University of Tokyo(東京大学

リチウムイオン二次電池正極／電解質界面構造の解明と設計

京都大学0048新制・課程博士博士(人間・環境学)甲第19072号人博第725号新制||人||174(附属図書館)26||人博||725(吉田南総合図書館)32023京都大学大学院人間・環境学研究科相関環境学専攻(主査)教授 内本 喜晴, 教授 加藤 立久, 教授 吉田 寿雄学位規則第4条第1項該当Doctor of Human and Environmental StudiesKyoto UniversityDGA

Phospholamban Ablation Using CRISPR/Cas9 System Improves Mortality in a Murine Heart Failure Model

<div><p>Sarcoplasmic reticulum Ca²⁺-ATPase 2a (SERCA2a) and its inhibitory protein called phospholamban (PLN) are pivotal for Ca²⁺ handling in cardiomyocyte and are known that their expression level and activity were changed in the heart failure patients. To examine whether PLN inhibition can improve survival rate as well as cardiac function in heart failure, we performed PLN ablation in calsequestrin overexpressing (CSQ-Tg) mice, a severe heart failure model, using clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated (Cas) system. According this method, generation rate of PLN wild type mice (PLN copy >0.95) and PLN homozygous knockout (KO) mice (PLN copy <0.05) were 39.1% and 10.5%, respectively. While CSQ overexpression causes severe heart failure symptoms and premature death, a significant ameliorating effect on survival rate was observed in PLN homozygous KO/CSQ-Tg mice compared to PLN wild type/CSQ-Tg mice (median survival days are 55 and 50 days, respectively). Measurement of cardiac function with cardiac catheterization at the age of 5 weeks revealed that PLN ablation improved cardiac function in CSQ-Tg mice without affecting heart rate and blood pressure. Furthermore, increases in atrial and lung weight, an index of congestion, were significantly inhibited by PLN ablation. These results suggest that PLN deletion would be a promising approach to improve both mortality and cardiac function in the heart failure.</p></div

Hemodynamic parameters.

<p>Cardiac catheterization was performed in CSQ non-Tg mice, PLN wild type (WT)/CSQ-Tg mice, and PLN homozygous KO/CSQ-Tg mice. (A) dP/dt_max, (B) dP/dt_min, (C) Tau, (D) LVEDP, (E) MBP, (F) HR. Values represent the mean ± SD, *P < 0.05, **P < 0.01 vs. CSQ non-Tg mice, ^#P < 0.05 vs. PLN WT/CSQ-Tg mice by Student's <i>t</i>-test.</p

Number of transferred embryos, pups, PLN homozygous KO mice, and PLN wild type (WT) mice in five lots.

<p>Number of transferred embryos, pups, PLN homozygous KO mice, and PLN wild type (WT) mice in five lots.</p

mRNA levels in the left ventricle.

<p>Analysis of mRNA levels was performed in the left ventricle of CSQ non-Tg mice, PLN wild type (WT)/CSQ-Tg mice, and PLN homozygous KO/CSQ-Tg mice. (A) β-MHC, (B) ANF, (C) BNF. Values represent the mean ± SD, *P < 0.05, **P < 0.01 vs. CSQ non-Tg mice, ^#P < 0.05 vs. PLN WT/CSQ-Tg mice by Student's <i>t</i>-test.</p

Number of CSQ-Tg and Non-Tg mice in five lots.

<p>Number of CSQ-Tg and Non-Tg mice in five lots.</p

sgRNAs and ssODN targeting <i>Pln</i> coding region.

<p>Targeting sequences of each sgRNA are capitalized and NGG PAM (protospacer-adjacent motif) sequences are underlined. Exons are indicated by closed boxes. The sgRNA targeting sites were designed to sandwich <i>Pln</i> coding region (filled with black). The ssODN is containing homologies of 60 bases on both sides flanking each of the sgRNA targeting sequences.</p