JP6189381B2 - 標的細胞を光刺激するためのシステム、方法、および組成物 - Google Patents
標的細胞を光刺激するためのシステム、方法、および組成物 Download PDFInfo
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Description
本特許明細書は、「標的細胞を光刺激するためのシステム、方法、および組成物」というタイトルの2008年4月23日に提出された米国仮特許出願第61/047,219号の米国特許法第119条(e)に基づく利益を主張する。この基礎となる仮出願全体を本明細書に参照により取り込む。
2009年4月7日に作成された「STFD_212PCT_ST25.txt」というファイル名の、8,345バイトのASCII(テキスト)ファイルである、本明細書と同時に提出されるコンピュータで読取り可能なヌクレオチド/アミノ酸配列リストの全体を参照により組み込む。
(1) 光刺激に反応する光活性化型イオンチャネル分子の使用方法であって、
細胞中に前記光活性化型イオンチャネル分子を設計付与すること;および
前記イオンチャネル分子に与えられ且つChR2イオンチャネルを活性化しない特性を有する光刺激に反応して、前記イオンチャネル分子を活性化し、イオンに前記光活性化型イオンチャネル分子を通過させることを含む、方法。
(2)前記光刺激の特性が波長および強度である、(1)に記載の方法。
(3)前記イオンチャネル分子が単一タンパク質コンポーネントである、(1)に記載の方法。
(4)前記細胞が幹細胞であり、前記活性化工程が、前記細胞の運命を制御する目的で行われる、(1)に記載の方法。
(5)前記活性化工程が関わる治療的効果を評価する工程を更に含み、前記評価が、前記細胞のモニタリング特性の関数として行われる、(1)に記載の方法。
(6)前記活性化工程の前後の前記細胞の特性をモニターし比較することにより、薬物に影響を与えるイオンチャネルをスクリーニングする工程を更に含む、(1)に記載の方法。
(7)前記光刺激を送達するために人工装具デバイスを追加する工程を更に含み、前記光刺激の波長が約530nmより大きい、(1)に記載の方法。
(8)細胞中に第2の光活性化型イオンチャネル分子を設計付与する工程を更に含み、前記第2の光活性化型イオンチャネル分子が約530nmより大きい波長の光刺激に反応しない、(1)に記載の方法。
(9)前記光活性化型イオンチャネル分子を用いて2つのニューロン集団の少なくとも1つでスパイクを発生させることによって前記2つのニューロン集団間で長期増強および長期抑圧の一方を促進する工程を更に含む、(1)に記載の方法。
(10)前記光刺激が一連の光パルスを含み、各パルスの長さが、各光パルスに対して個々の活動電位を起こさせるのに十分である、(1)に記載の方法。
(11)前記光活性化型イオンチャネル分子を通過する前記イオンにナトリウムイオンが含まれる、(1)に記載の方法。
(12)前記設計付与工程が、前記光活性化型イオンチャネル分子を含むウイルスベクターを用いた前記細胞の形質導入、前記細胞のトランスフェクション、前記細胞へのDNAのマイクロインジェクション、および特定のクラスの細胞への前記光活性化型イオンチャネル分子の遺伝的ターゲティングの少なくとも1つを含む、(1)に記載の方法。
(13)前記細胞中の生じたイオン濃度を光センサーを用いて測定する工程を更に含む、(1)に記載の方法。
(14)前記活性化工程が、外因性補因子を用いない、(1)に記載の方法。
(15)前記光活性化型イオンチャネル分子が、ボルボックス(Volvox carteri)に由来する遺伝子から発現される、(1)に記載の方法。
(16)約530nmより大きい波長の光が当たると反応して活性化する光活性化型イオンチャネルを有する分子を発現するためのヌクレオチド配列。
(17)細胞中で異種(heterologous)分子を発現するためのヌクレオチド配列であって、前記分子が、約530nmより大きい波長の光が当たると反応して活性化する光活性化型イオンチャネル分子を有する、ヌクレオチド配列。
(18)細胞中で異種(heterologous)分子を発現するためのヌクレオチド配列を含むプラスミドであって、前記分子が、約530nmより大きい波長の光が当たると反応して活性化する光活性化型イオンチャネルを有する、プラスミド。
(19)細胞中で異種(heterologous)光活性化型イオンチャネル分子を発現する細胞であって、前記分子が、約530nmより大きい波長の光に反応して前記イオンチャネルを活性化する、細胞。
(20)配列番号3に示すタンパク質を異種(heterologous)発現するためのヌクレオチド配列。
(21)配列番号3に示す異種(heterologous)タンパク質を発現するための細胞。
(22)2種類の異種(heterologous)光活性化型イオンチャネル分子を発現する細胞であって、前記分子が、各イオンチャネルの独立制御ができるようにそれぞれの波長の光に反応して前記イオンチャネルを活性化する、細胞。
(23)パーキンソン病の治療方法であって、
視床下核(STN)の求心性軸索の局所的脱分極を与えるために刺激デバイスを配置する工程;および
前記刺激デバイスおよび刺激プロファイルを利用して前記求心性軸索を直接脱分極する工程を含む方法。
(24)求心性軸索中に分子を設計付与する工程を更に含み、前記分子が光に反応してイオンチャネルを活性化する、(23)に記載の方法。
(25)前記刺激デバイスが光送達デバイスである、(24)に記載の方法。
(26)前記刺激プロファイルが約90〜130Hzである、(24)に記載の方法。
(27)前記刺激プロファイルが約20Hzである、(24)に記載の方法。
(28)前記求心性軸索の局所的脱分極が、一次運動皮質の求心性軸索を標的とする、(24)に記載の方法。
(29)光に反応して活性化するイオンチャネルを有する分子を発現し一次運動皮質の求心性軸索に標的化される遺伝子配列を設計付与する工程を更に含む、(23)に記載の方法。
Claims (14)
- a)発光器、及び
b)ボルボックス(Volvox carteri)の光反応性イオンチャネルタンパク質(VChR1)を発現するヌクレオチド配列を有する核酸を含む生物学的配置体、を備え、
前記光反応性イオンチャネルタンパク質VChR1が535nm〜589nmの範囲の波長を持つ光で活性化され、
前記ヌクレオチド配列が哺乳動物細胞での発現のために最適化される、埋め込み可能なデバイス。 - 前記光反応性イオンチャネルタンパク質VChR1が、配列番号:3に示されるアミノ酸配列を有する、請求項1に記載のデバイス。
- 前記核酸が発現ベクターである、請求項1に記載のデバイス。
- 前記発現ベクターがレンチウイルスベクターである、請求項3に記載のデバイス。
- 前記発現ベクターがアデノ随伴ウイルスベクターである、請求項3に記載のデバイス。
- 前記ヌクレオチド配列が特定のニューロン集団での発現を誘導するプロモータに作動可能に結合する、請求項1に記載のデバイス。
- 前記ヌクレオチド配列がアルファ−CaMKIIレンチウイルスプロモータに作動可能に結合する、請求項6に記載のデバイス。
- 前記発光器が発光ダイオードを含む、請求項1に記載のデバイス。
- 前記発光器が光ファイバーーを含む、請求項1に記載のデバイス。
- 前記発光器がレーザーを含む、請求項1に記載のデバイス。
- 前記生物学的配置体が、クラミドモナス(Chlamydomonas reinhardtii)由来の光反応性タンパク質ChR2を発現するヌクレオチド配列を有する核酸を含む、請求項1に記載のデバイス。
- 前記光反応性タンパク質ChR2が、配列番号:2に示されるアミノ酸配列を有する、請求項11に記載のデバイス。
- 前記光反応性タンパク質ChR2が、589nmの波長を持つ光で活性化される、請求項11に記載のデバイス。
- 前記生物学的配置体が、光反応性タンパク質NpHRを発現するヌクレオチド配列を有する核酸を含む、請求項1に記載のデバイス。
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