JP6141517B2 - 脂肪細胞標的非ウイルス性遺伝子伝達体 - Google Patents
脂肪細胞標的非ウイルス性遺伝子伝達体 Download PDFInfo
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Description
DMEM(Dulbecco’s Modified Eagle Medium)、グルコース、FBSをWelGENE(韓国)から購入し、インスリン及び及び3―isobutyl―1―methylxanthine(IBMX)をWako(日本)から、デキサメタゾン及びOil Red OをSigma―Aldrich(米国)から購入した。3T3―L1前駆脂肪細胞(preadipocyte)を韓国細胞株銀行から購入し、その分化を誘導した。
ローダミンB結合されたATS(CKGGRAKDC)ペプチド、FITC結合されたATS―R9(CKGGRAKDRRRRRRRRRC)及びR9(CRRRRRRRRRC)ペプチドをPeptron(Daejoen、Korea)社から入手した。凍結・乾燥したペプチドを脱イオン水に溶かした後、使用前まで−20℃で保管した。
脂肪細胞分化をOil Red O染色で確認した。
Cellmask Deep RedをInvitrogenから購入し、DAPI―Fluoromount―GをSouthern Biotechから購入した。3T3―L1前駆脂肪細胞、H9c2及びHEK293細胞を6ウェルプレートに置いたカバースリップ上で成長及び分化させた。
競争分析(competition assay)のために、3T3―L1前駆脂肪細胞と成熟脂肪細胞(10日目)に対して自由ATS(free―ATS)(100μg/mL)を処理した。培養してから6時間後、各細胞をFITC―ATS―R9オリゴ―ペプトプレックスで処理して24時間培養した。H9c2及びHEK293細胞株を対照群として使用した。H9c2及びHEK293細胞に対してFITC結合されたATS―R9(25μg/mL)オリゴ―ペプトプレックスを処理し、これを24時間培養した。
1μgのルシフェラーゼプラスミドDNA(p―β―Luci、Promega(USA)から入手)、脱イオン水及びATS―R9(電荷比率:0.25、0.5、1、3、5、7、及び9)を30分間常温で培養してオリゴペプトプレックス(oligo―peptoplexes)を製作し、0.8%のアガロースゲルで0.5%のTBE緩衝溶液下で25分間電気泳動を行った(100V)。
5μgのルシフェラーゼプラスミドDNA(p―β―Luci)、脱イオン水及びATS―R9(電荷比率:0.5、1、3、5、7、及び9)を30分間常温で培養し、オリゴペプトプレックス(oligo―peptoplexes)を製作した。前記オリゴペプトプレックスの平均直径及び表面ゼータポテンシャルは、Zetasizer―Nano ZS(Malvern Instruments、UK)を備えたDLSを使用して測定した。
ルシフェラーゼ分析キットをPromega(USA)から購入し、DCタンパク質分析キット及び小血清アルブミンスタンダードをBio―Rad Laboratories(USA)から購入した。前駆脂肪細胞3T3―L1及び分化した脂肪細胞を24ウェルプレートにシーディングした。
細胞生存能をMTT[3―(4,5―dimethylthiazol―2―yl)―2,5―diphenyltetrazolium bromide]分析法で測定した。
3週齢のC57BL/6JマウスをCentral Lab Animal Inc.(Korea)から購入した。最初の2週間は、50%の正常食餌及び脂肪から60kcal%のローデント食餌(rodent diet)で飼育した。マウスに対する高脂肪食餌比率を漸次増加させ、7週に開始した;前記各マウスを脂肪から60kcal%で高脂肪食餌でのみ飼育した。14週後、各マウスは肥満になった(体重>45g)。
FITC結合されたATS―R9(10mg/mL)及びFITC―R9(10mg/mL)をPBSに希釈し、最終濃度を1.5mg/mLとし、肥満マウスに尾静脈注射で100μlのペプチドを注入した。
3T3―L1脂肪細胞分化過程を図2に示した。
ATS―R9の細胞内への内在化程度をモニターするために、FITC―標識されたATS―R9(FITC―ATS―R9)で成熟脂肪細胞及び前駆脂肪細胞を処理し、担体の細胞内伝達を確認した。
ATS―R9の脂肪細胞ターゲッティングに対するATSの影響を確認するために競争分析を行った。
脂肪細胞の分化程度によるプロヒビチン発現の位置及び量を確認するために、二つのタイプの脂肪細胞をローダミンB―標識されたATS(RhoB―ATS)で処理した。ATSの高いプロヒビチン結合能力のため、プロヒビチンを確認するための抗体の代わりにATSを使用することができる。
本発明の融合オリゴ―ペプトプレックスが肥満脂肪細胞特異的な特性を有していることをより明確に確認するために、非―肥満マウスにおけるプロヒビチン発現様相と、筋肉細胞におけるプロヒビチン発現様相とを比較して分析した。このとき、ウエスタンブロット方法を用いた。そして、その結果を図6に示した。
陽イオン性ポリマー/DNA複合体の生化学的特性は、細胞吸収能、カーゴ(cargo)安定性、細胞毒性及びトランス遺伝子発現に影響を及ぼす主要な要素の一つである。ATS―R9のDNA凝縮及び保護能力をゲル遅延分析及びマウス血清における分解(degradation)テストによって試験した。
ATS―R9の凝縮効能を分枝型PEI(25kDa)の場合と比較した。
次に、ATS―R9/DNAオリゴ―ペプトプレックスのゼータポテンシャル及び平均直径をDLS(dynamic light scattering)を用いて測定した。
また、ATS―R9/DNAオリゴ―ペプトプレックスの毒性を試験した。10日目に脂肪細胞に分化した3T3―L1にオリゴ―ペプトプレックスを形質転換した。48時間後、細胞生存能をMTT分析で測定した(n=6)。
6―1.最適な電荷比率
トランスフェクション効率の最適な電荷比率を検討した。
そして、トランスフェクションの最適時期を調査するために、3T3―L1分化した各脂肪細胞を異なる分化日付にルシフェラーゼプラスミドDNA(p―β―Luci)で電荷比率5でATS―R9及びPEIによってトランスフェクションした。ルシフェラーゼ遺伝子発現は、ルシフェラーゼ分析キットによって72時間後に測定した。
食餌誘導肥満(diet―induced obesity、DIO)マウスを用いてin vivo実験を行った。脂肪組織に対するATS―R9のターゲッティングを観察するために、FITC―ATS―R9を肥満マウスに尾静脈を通じて投与した。プローブ基盤の共焦点レーザーエンドマイクロスコピー(pCLE、Cellvizio)を用いて追跡を可視化した。
まず、in vivo上の脂肪組織内にATS―R9が内在化することを確認した。
C57BL/6J肥満マウスに尾静脈を通じてFITC―コンジュゲートされたATS―R9を注入し、Rhodamine―コンジュゲートされたレクチンを注入することによって血管染色を行った。
本発明のATS―R9がin vitro及びin vivoでsh―RNA伝達と脂肪細胞へのサイレント遺伝子伝達において優れた効果を有するか否かを確認しようとした。
また、sh―FABP4処理後、体重減少も確認された(図16)。
また、インスリン及びグルコース耐性を観察した。
(1)他の伝達体の脂肪細胞への遺伝子伝達能
PEIとlipofectaminを比較群とし、肥満脂肪細胞への遺伝子luciferase伝達能力を評価し、これを図19に示した。
脂肪細胞の分化前と分化後の遺伝子伝達効能を比較した結果を図20に示した。
伝達遺伝子としてluciferaseを使用し、脂肪ターゲッティング配列としてScrambled ATSを使用して肥満脂肪細胞への遺伝子伝達効能を比較した。
その結果、図21で確認できるように、ATSの配列をスクランブルして伝達した場合、遺伝子伝達能力が確実に低下することを確認することができた。
Claims (15)
- 配列番号1で表示されるアミノ酸配列からなる脂肪細胞標的配列(ATS)及びR9(arginine)ペプチドを含有する脂肪細胞標的遺伝子伝達体。
- 前記脂肪細胞標的配列及びR9(arginine)ペプチドは、プロヒビチン(prohibitin)と結合されることを特徴とする、請求項1に記載の脂肪細胞標的遺伝子伝達体。
- 前記R9(arginine)ペプチドは、Cys―(D―R)9―Cys構造を有することを特徴とする、請求項1又は2に記載の脂肪細胞標的遺伝子伝達体。
- 前記脂肪細胞は、分化した成熟肥満脂肪細胞であることを特徴とする、請求項1〜3のいずれか1項に記載の脂肪細胞標的遺伝子伝達体。
- 前記脂肪細胞は、分化してから9日〜11日経過した成熟した肥満脂肪細胞であることを特徴とする、請求項4に記載の脂肪細胞標的遺伝子伝達体。
- 配列番号1で表示されるアミノ酸配列からなる脂肪細胞標的配列(ATS)、R9(arginine)ペプチド、及び脂肪細胞標的遺伝子として肥満及び肥満由来代謝症候群治療遺伝子を含有する複合体。
- 前記脂肪細胞標的配列及びR9(arginine)ペプチドは、プロヒビチン(prohibitin)と結合されることを特徴とする、請求項6に記載の複合体。
- 前記R9(arginine)ペプチドは、Cys―(D―R)9―Cys構造を有することを特徴とする、請求項6又は7に記載の複合体。
- 前記肥満及び肥満由来代謝症候群治療遺伝子は、DNA又はRNAiであることを特徴とする、請求項6〜8のいずれか1項に記載の複合体。
- 前記RNAiは、siRNA又はshRNAであることを特徴とする、請求項9に記載の複合体。
- 前記複合体は、200nm以下の直径を有することを特徴とする、請求項6〜10のいずれか1項に記載の複合体。
- 前記複合体は、3:1〜8:1の電荷比率(+/−)を有することを特徴とする、請求項6〜11のいずれか1項に記載の複合体。
- 前記脂肪細胞は、分化した成熟肥満脂肪細胞であることを特徴とする、請求項6〜12のいずれか1項に記載の複合体。
- 前記脂肪細胞は、分化してから9日〜11日経過した成熟した肥満脂肪細胞であることを特徴とする、請求項13に記載の複合体。
- 請求項6〜14のいずれか1項に記載の複合体を有効成分として含有する肥満又は肥満由来代謝症候群治療用組成物。
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CA3163942A1 (en) * | 2020-01-15 | 2021-07-22 | Kuen Yong Lee | Gas-generating micelle for reducing localized fat |
CN111249468B (zh) * | 2020-03-18 | 2022-06-24 | 湖南大学 | 一种脂肪细胞靶向的dna纳米药物及其制备方法与应用 |
CN111394392B (zh) * | 2020-03-24 | 2022-03-29 | 中国科学院长春应用化学研究所 | 一种脂肪细胞靶向阳离子基因载体、其制备方法及其应用 |
CN114288250B (zh) * | 2022-03-10 | 2022-06-03 | 北京同柏宸科生物技术开发有限公司 | 一种脂肪细胞靶向的跨膜转运脂质体药物载体及其制备方法和用途 |
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Family Cites Families (15)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP0975370B9 (en) | 1997-05-21 | 2004-11-03 | The Board Of Trustees Of The Leland Stanford Junior University | Composition and method for enhancing transport across biological membranes |
EP1378515A1 (en) * | 2002-07-01 | 2004-01-07 | Deutsches Krebsforschungszentrum Stiftung des öffentlichen Rechts | Peptides for inducing apoptosis in tumor cells |
US20060024692A1 (en) | 2002-09-30 | 2006-02-02 | Oncotherapy Science, Inc. | Method for diagnosing non-small cell lung cancers |
TW200413725A (en) * | 2002-09-30 | 2004-08-01 | Oncotherapy Science Inc | Method for diagnosing non-small cell lung cancers |
CA2513072A1 (en) | 2003-01-09 | 2004-07-29 | Invitrogen Corporation | Cellular delivery and activation polypeptide-nucleic acid complexes |
US20060122118A1 (en) | 2004-12-03 | 2006-06-08 | Ho Siew P | Therapeutic compositions and methods of using same |
AU2007314366A1 (en) * | 2006-10-30 | 2008-05-08 | Southern Research Institute | Targeting NBS1-ATM interaction to sensitize cancer cells to radiotherapy and chemotherapy |
MX360094B (es) * | 2007-06-19 | 2018-10-19 | Kythera Biopharmaceuticals Inc Star | Composición, método y preparación del ácido biliar sintético. |
CN101801340A (zh) * | 2007-06-28 | 2010-08-11 | 新加坡科技研究局 | 用于将制剂递送到细胞中的阳离子肽 |
KR101035364B1 (ko) | 2008-05-21 | 2011-05-20 | 한양대학교 산학협력단 | 핵산 전달용 세포내 환원성 폴리(올리고-아르기닌) |
CN102120035A (zh) * | 2010-05-24 | 2011-07-13 | 江苏省人民医院 | 一种靶向至白色脂肪组织治疗代谢综合征的新型靶向药物 |
CN103221070B (zh) | 2010-08-30 | 2019-07-12 | 哈佛大学校长及研究员协会 | 用于狭窄病变和溶解血栓疗法的切变控制释放 |
CN102010461B (zh) * | 2010-10-11 | 2014-02-12 | 华南理工大学 | 一种alpha螺旋状阳离子多肽分子及其制法和应用 |
US20140056811A1 (en) * | 2010-12-27 | 2014-02-27 | Compugen Ltd. | New cell-penetrating peptides and uses thereof |
CN102174110B (zh) * | 2011-01-24 | 2013-06-05 | 西南大学 | 狂犬病毒糖蛋白衍生肽及其应用 |
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