National Institutes of Health/National Institute of Neurological Disorders and Stroke (NIH/NINDS)
DP1.NS111369
米国
National Institutes of Health/National Institute of Mental Health (NIH/NIMH)
UF1.MH128336
米国
引用
ジャーナル: Nat Commun / 年: 2024 タイトル: Human cell surface-AAV interactomes identify LRP6 as blood-brain barrier transcytosis receptor and immune cytokine IL3 as AAV9 binder. 著者: Timothy F Shay / Seongmin Jang / Tyler J Brittain / Xinhong Chen / Beth Walker / Claire Tebbutt / Yujie Fan / Damien A Wolfe / Cynthia M Arokiaraj / Erin E Sullivan / Xiaozhe Ding / Ting-Yu ...著者: Timothy F Shay / Seongmin Jang / Tyler J Brittain / Xinhong Chen / Beth Walker / Claire Tebbutt / Yujie Fan / Damien A Wolfe / Cynthia M Arokiaraj / Erin E Sullivan / Xiaozhe Ding / Ting-Yu Wang / Yaping Lei / Miguel R Chuapoco / Tsui-Fen Chou / Viviana Gradinaru / 要旨: Adeno-associated viruses (AAVs) are foundational gene delivery tools for basic science and clinical therapeutics. However, lack of mechanistic insight, especially for engineered vectors created by ...Adeno-associated viruses (AAVs) are foundational gene delivery tools for basic science and clinical therapeutics. However, lack of mechanistic insight, especially for engineered vectors created by directed evolution, can hamper their application. Here, we adapt an unbiased human cell microarray platform to determine the extracellular and cell surface interactomes of natural and engineered AAVs. We identify a naturally-evolved and serotype-specific interaction between the AAV9 capsid and human interleukin 3 (IL3), with possible roles in host immune modulation, as well as lab-evolved low-density lipoprotein receptor-related protein 6 (LRP6) interactions specific to engineered capsids with enhanced blood-brain barrier crossing in non-human primates after intravenous administration. The unbiased cell microarray screening approach also allows us to identify off-target tissue binding interactions of engineered brain-enriched AAV capsids that may inform vectors' peripheral organ tropism and side effects. Our cryo-electron tomography and AlphaFold modeling of capsid-interactor complexes reveal LRP6 and IL3 binding sites. These results allow confident application of engineered AAVs in diverse organisms and unlock future target-informed engineering of improved viral and non-viral vectors for non-invasive therapeutic delivery to the brain.
全体 : Adeno-associated virus-9 and human interleukin 3 complex
全体
名称: Adeno-associated virus-9 and human interleukin 3 complex
要素
複合体: Adeno-associated virus-9 and human interleukin 3 complex
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超分子 #1: Adeno-associated virus-9 and human interleukin 3 complex
超分子
名称: Adeno-associated virus-9 and human interleukin 3 complex タイプ: complex / ID: 1 / 親要素: 0 / 含まれる分子: #1-#2 詳細: A complex of adeno-associated virus-9 and human Interleukin 3. Both proteins were purified from a human cell-based expression system.
由来(天然)
生物種: Adeno-associated virus 9 (アデノ随伴ウイルス)
分子量
理論値: 46.8 KDa
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実験情報
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構造解析
手法
クライオ電子顕微鏡法
解析
サブトモグラム平均法
試料の集合状態
particle
-
試料調製
緩衝液
pH: 7.6 / 構成要素:
濃度
名称
20.0 mM
PBS
0.001 %
Pluronic-F68
凍結
凍結剤: ETHANE / チャンバー内湿度: 100 % / チャンバー内温度: 293 K / 装置: FEI VITROBOT MARK IV