A novel bone-targeting AAV-mediated gene therapy to promote bone formation in osteoporosis

一种新型骨靶向 AAV 介导的基因疗法可促进骨质疏松症的骨形成

基本信息

批准号：
10458098
负责人：
Jae-Hyuck Shim
金额：
$ 21.89万
依托单位：
UNIV OF MASSACHUSETTS MED SCH WORCESTER
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-08-01 至 2024-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10458098
关键词：
Adaptor Signaling Protein Adult Adverse effects Affinity Chromatography Age Aging American Anabolic Agents Angiogenic Factor Antibodies Binding Sites Biodistribution Biological Bone Density Bone Resorption Bone necrosis Bone structure Candidate Disease Gene Capsid Cell Lineage Cells Clinical Data Deterioration Disease Endothelium Engineering Equilibrium Estrogens FDA approved Femoral Fractures Gene Transduction Agent Gene Transfer Health Histology Histopathology In Vitro Infection Injections Intravenous Jaw Label Liver Mass Spectrum Analysis Measures Mediating MicroRNAs Modification Mus Myocardium Operative Surgical Procedures Osteoblasts Osteoclasts Osteocytes Osteogenesis Osteoporosis Osteoporotic Ovariectomy PTH gene Pathogenicity Pathway interactions Patients Phenotype Postmenopausal Osteoporosis Proteins Proteomics RNA Interference Recombinant adeno-associated virus (rAAV)Repression Reverse Transcriptase Polymerase Chain Reaction Senile Osteoporosis Serotyping Serum Calcium Level Skeletal Muscle Therapeutic Therapeutic Agents Therapeutic Intervention Tissues Tropism Validation Viral Vector WNT Signaling Pathway aged antagonist base biomechanical test bone bone aging bone loss bone mass calcification fracture risk gene therapy immunogenicity improved in vivo innovation knock-down long bone mechanical properties mouse model new therapeutic target novel optical imaging osteogenic osteoporosis with pathological fracture parathyroid hormone-related protein response side effect skeletal spine bone structure targeted treatment therapeutic gene therapeutic target transcriptome transduction efficiency transgene expression vector

项目摘要

PROJECT SUMMARY Adult bone mass is determined by the balance between bone formation by osteoblasts (OBs) and bone resorption by osteoclasts (OCs), and disturbances in this equilibrium to favor osteoclast (OC)-mediated resorption leads to osteoporosis. The majority of existing therapeutics for osteoporosis act by inhibiting OCs, but these can not cure osteoporosis and are limited by rare side effects. Current anabolic agents, parathyroid hormone (PTH), parathyroid hormone-related protein (PTHrP), and anti-sclerostin antibody exist for promoting osteoblast (OB) function to treat patients with osteoporosis. However, these agents are also limited by concern for off-target adverse effects and waning efficacy. Previously, we and others demonstrated that inhibition of potent OB suppressors, sclerostin (SOST) and the adaptor protein schnurri-3 (SHN3), promotes bone formation in mouse models of postmenopausal and senile osteoporosis. One innovative approach to treat osteoporosis is RNAi-based bone anabolic gene therapy using recombinant adeno-associated virus (rAAV). Using our engineered rAAV9 vector with bone-specific tropism and transgene expression, we will develop novel gene therapeutics that promote bone formation in osteoporosis with a single systemic administration. Additionally, we will identify potential novel osteogenic and/or angiogenic factors regulated by the SOST/SHN3 pathway that could be useful as therapeutic targets for osteoporosis. Aim 1 will examine whether bone-specific rAAV9-mediated silencing of SHN3 or SOST can reverse bone loss in mouse models of osteoporosis. To avoid potential off-target adverse effects in non-skeletal tissues, rAAV9’s bone- specific tropism and transgene expression were further improved by capsid modification and tissue-specific miRNA-mediated repression of rAAV expression. Using two mouse models of osteoporosis (ovariectomized (OVX) and aged mice), we will determine therapeutic potentials of rAAV9-mediated silencing of SHN3 or SOST in postmenopausal and senile osteoporosis. Aim 2 will identify novel osteogenic and/or angiogenic factors regulated by the SHN3/SOST pathway that promote bone formation in osteoporosis. In vivo silencing accuracy of rAAV9 carrying amiR-shn3 or amiR-sost in bone-residing OB lineage cells will be examined by scatterplot analysis of whole transcriptome data. Once validated, wt, shn3- or sost-deficient OB-lineage cells will be FACS-sorted from AAV-treated, OVX-mice and subjected for transcriptome profiling. As a complimentary approach, proteomics will be performed in AAV-transduced OB-lineage cells isolated from MetRS;Prx1-cre mice with OB-specific labeling of nascent proteins after OVX-surgery. These combinatory approaches will allow us to identify novel osteogenic and/or angiogenic factors commonly or differentially regulated by the pathways of SHN3 and SOST in response to estrogen deficiency-induced osteoporosis. Successful completion of these aims will provide proof-of-concept demonstration and identify potential novel regulators that could be useful as therapeutic targets for osteoporosis.

项目概要成人骨量由成骨细胞（OB）形成的骨和骨之间的平衡决定破骨细胞（OC）的吸收，以及这种平衡的紊乱有利于破骨细胞（OC）介导吸收导致骨质疏松症大多数现有的骨质疏松症治疗方法是通过抑制 OCs 起作用。但这些药物不能治愈骨质疏松症，并且受到当前合成代谢药物甲状旁腺罕见副作用的限制。激素（PTH）、甲状旁腺激素相关蛋白（PTHrP）和抗硬化素抗体的存在可促进成骨细胞（OB）的功能可以治疗骨质疏松症患者，但是，这些药物也受到限制。此前，我们和其他人证明了对脱靶副作用和疗效减弱的抑制作用。强效 OB 抑制剂、硬化蛋白 (SOST) 和接头蛋白 schnurri-3 (SHN3)，可促进骨生长一种治疗绝经后和老年骨质疏松症的创新方法。骨质疏松症是使用重组腺相关病毒进行基于 RNAi 的骨合成代谢基因治疗 (rAAV)。使用我们的具有骨特异性向性和转基因表达的工程化 rAAV9 载体，我们将开发新的基因疗法，通过单一系统促进骨质疏松症的骨形成此外，我们将鉴定潜在的新型成骨和/或血管生成因子。目标 1 将研究可作为骨质疏松症治疗靶点的 SOST/SHN3 通路。骨特异性 rAAV9 介导的 SHN3 或 SOST 沉默是否可以逆转小鼠骨质流失为了避免非骨骼组织中潜在的脱靶副作用，rAAV9 的骨- 通过衣壳修饰和组织特异性进一步改善了特异性向性和转基因表达使用两种骨质疏松症小鼠模型（卵巢切除）的 miRNA 介导的 rAAV 表达抑制。（OVX）和老年小鼠），我们将确定 rAAV9 介导的 SHN3 或 SOST 沉默的治疗潜力目标 2 将确定新的成骨和/或血管生成因子。受 SHN3/SOST 通路调节，促进骨质疏松症中的骨形成。将通过以下方法检查骨驻留 OB 谱系细胞中携带 amiR-shn3 或 amiR-sost 的 rAAV9 的准确性验证后，对 wt、shn3 或 sost 缺陷的 OB 谱系细胞进行全转录组数据的散点图分析。将从 AAV 处理的 OVX 小鼠中进行 FACS 分选，并进行转录组分析。补充方法，蛋白质组学将在 AAV 转导的 OB 谱系细胞中进行，其中分离自 MetRS;Prx1-cre 小鼠在 OVX 手术后具有 OB 特异性标记的新生蛋白。方法将使我们能够共同或差异地识别新的成骨和/或血管生成因子通过 SHN3 和 SOST 通路调节雌激素缺乏引起的骨质疏松症。成功完成这些目标将提供概念验证演示并确定潜在的新型调节剂可用作骨质疏松症的治疗靶点。

项目成果

期刊论文数量（5）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Impaired mitochondrial oxidative metabolism in skeletal progenitor cells leads to musculoskeletal disintegration.

骨骼祖细胞中线粒体氧化代谢受损导致肌肉骨骼解体。

DOI：
发表时间：
2022-11-11
期刊：
影响因子：
16.6
作者：
Lin, Chujiao;Yang, Qiyuan;Guo, Dongsheng;Xie, Jun;Yang, Yeon;Chaugule, Sachin;DeSouza, Ngoc;Oh, Won;Li, Rui;Chen, Zhihao;John, Aijaz A;Qiu, Qiang;Zhu, Lihua Julie;Greenblatt, Matthew B;Ghosh, Sankar;Li, Shaoguang;Gao, Guangping;H
通讯作者：
H

Gene Therapy for Fibrodysplasia Ossificans Progressiva: Feasibility and Obstacles.

进行性骨化性纤维发育不良的基因治疗：可行性和障碍。

DOI：
发表时间：
2022-08
期刊：
影响因子：
4.2
作者：
Eekhoff, Elisabeth M W;de Ruiter, Ruben D;Smilde, Bernard J;Schoenmaker, Ton;de Vries, Teun J;Netelenbos, Coen;Hsiao, Edward C;Scott, Christiaan;Haga, Nobuhiko;Grunwald, Zvi;De Cunto, Carmen L;di Rocco, Maja;Delai, Patricia L R;Diecidue, Robe
通讯作者：
Diecidue, Robe

AAV-mediated delivery of osteoblast/osteoclast-regulating miRNAs for osteoporosis therapy.

AAV 介导的成骨细胞/破骨细胞调节 miRNA 的递送用于骨质疏松症治疗。

DOI：
发表时间：
2022-09-13
期刊：
影响因子：
0
作者：
John, Aijaz Ahmad;Xie, Jun;Yang, Yeon;Kim, Jung;Lin, Chujiao;Ma, Hong;Gao, Guangping;Shim, Jae
通讯作者：
Shim, Jae

Regulation of sclerostin by the SIRT1 stabilization pathway in osteocytes.

骨细胞中 SIRT1 稳定途径对硬化素的调节。

DOI：
发表时间：
2022-08
期刊：
影响因子：
12.4
作者：
Kim, Jung;Yang, Yeon;Xie, Jun;Lee, Oksun;Kim, JiHea;Hong, Jaehyoung;Boldyreff, Brigitte;Filhol, Odile;Chun, Hyonho;Greenblatt, Matthew B;Gao, Guangping;Shim, Jae
通讯作者：
Shim, Jae

Biphasic regulation of osteoblast development via the ERK MAPK-mTOR pathway.

通过 ERK MAPK-mTOR 途径对成骨细胞发育进行双相调节。

DOI：
发表时间：
2022-08-17
期刊：
影响因子：
7.7
作者：
Kim, Jung;Yang, Yeon;Hong, Jaehyoung;Chaugule, Sachin;Chun, Hyonho;van der Meulen, Marjolein C H;Xu, Ren;Greenblatt, Matthew B;Shim, Jae
通讯作者：
Shim, Jae