Multiplex Engineered Human Lymphocytes for Therapeutic Protein Delivery

用于治疗性蛋白质递送的多重工程人类淋巴细胞

基本信息

批准号：
10447169
负责人：
Beau Richard Webber
金额：
$ 23.25万
依托单位：
UNIVERSITY OF MINNESOTA
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：
2021-07-07 至 2023-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10447169
关键词：
Address Allogenic Architecture Area Autologous Bulla COL7A1 CRISPR/Cas technology Cell Therapy Cell Transplantation Cell surface Cells Chronic Collagen Complementary DNA Deposition Dermal Disease Engineering Epidermolysis Bullosa Dystrophica Exhibits FDA approved Failure Fibroblasts GPR2 gene Gene Mutation Gene-Modified Genes Genetic Diseases Genome Genome engineering Goals Health Hematopoietic stem cells Hereditary Disease Histopathology Home Homing Human Human Engineering Immune Impairment In Vitro Inflammation Mediators Inflammatory Infusion procedures Inherited Injections Intervention Lymphocyte Mediating Membrane Methods Minnesota Modification Morbidity - disease rate Mucous Membrane Mus Mutation Nature Organ Palliative Care Pathologic Pathology Patients Phenotype Population Production Property Proteins Rare Diseases Reagent Recombinants Refractory Research Resolution Safety Skin Squamous cell carcinoma Stem cell transplant System T-Lymphocyte T-Lymphocyte Subsets Testing Therapeutic Therapeutic Effect Tissues Transcription Initiation Site Treatment Efficacy Universities Virus Visceral Zoledronate base base editor biomedical referral center cellular engineering comparative cytokine cytotoxic cytotoxicity design direct application engineered T cells gene function genetic referral center genome editing graft vs host disease hematopoietic repopulating cell immunodeficient mouse model improved in vivo innovation keratinocyte knockout gene mesenchymal stromal cell migration minimal risk mouse model new technology novel overexpression promoter public health relevance receptor residence stem cells tau Proteins therapeutic protein therapy development translational approach translational potential vector wound γδ T cells

项目摘要

Abstract. Recessive dystrophic epidermolysis bullosa (RDEB) is a severe autosomal recessive disease caused by collagen type VIIa (COL7A1) gene mutations. RDEB is characterized by absent/defective COL7A1 (C7) protein deposition causing severe blistering, mucosal tissue damage, and aggressive squamous cell carcinoma. The University of Minnesota serves as a leading treatment referral center for this inherited disorder. Palliative care is non-curative and cellular therapy options include autologous or allogeneic local and/or systemic infusion of keratinocytes, fibroblasts, mesenchymal stromal cells (MSC), or hematopoietic stem/progenitor cells (HSPC). None of these currently employed treatment options resolve the full pathological spectrum of RDEB. Active wound areas persist, and mucosal disease remains highly refractory to intervention contributing to significant morbidity. Keratinocytes and fibroblasts, the primary C7 producing cells, show limited migration and persistence following localized injection. MSC and HSPC have broad circulatory potential, however, they produce comparatively low levels of C7 and residence in the skin or mucosa is not well established. Thus, it is essential to develop more efficacious cellular therapies capable of accessing skin and mucocutaneous tissues. γδ T cells are abundant within skin and mucosa, and due to their MHC-unrestricted nature are compatible with allogeneic transfer, however they do not naturally produce C7. Our innovative approach will employ precision genome modification using CRISPR/Cas9 to engineer γδ T cells to produce high levels of endogenous C7. We hypothesize that the tissue migratory properties of γδ T cells—particularly to the skin and mucosa—as well as their demonstrated allo-compatibility, make them uniquely suited for therapeutic delivery of C7 protein. In Aim 1 we will define a genome engineering strategy to confer high C7 expression, enhanced skin homing, and reduced inflammatory/cytotoxic capacity to primary human γδ T cells. In Aim 2 we will evaluate the ability of engineered allogeneic γδ T cells to home to skin and mucosa, deposit C7, and ameliorate pathology in an immunodeficient mouse model of RDEB. Further, we will test the effect of zoledronate induced in vivo expansion of the engineered Vγ9Vδ2 T cell subset on therapeutic efficacy. Our approach is a highly novel and innovative allogeneic strategy designed to address key limitations of current cellular therapies for RDEB. The application of CRISPR/Cas9 in γδ T cells represents a novel engineering and protein delivery strategy with translational potential for RDEB, other inherited mucocutaneous disorders, and a multitude of diverse disorders treated by cell/stem cell transplant.

抽象的。隐性营养不良性大疱性表皮松解症（RDEB）是一种严重的常染色体隐性遗传病，由以下原因引起： VIIa 型胶原蛋白 (COL7A1) 基因突变的特点是 COL7A1 (C7) 蛋白缺失/缺陷。沉积导致严重起泡、粘膜组织损伤和侵袭性鳞状细胞癌。明尼苏达大学是这种遗传性疾病的领先治疗转诊中心。是非治愈性的，细胞治疗选择包括自体或同种异体局部和/或全身输注角质形成细胞、成纤维细胞、间充质基质细胞 (MSC) 或造血干/祖细胞 (HSPC)。目前使用的这些治疗方案均不能解决 RDEB Active 的全部病理谱。伤口区域持续存在，粘膜疾病仍然很难干预，导致严重的后果角质形成细胞和成纤维细胞（主要的 C7 产生细胞）显示出有限的迁移和持久性。局部注射后 MSC 和 HSPC 具有广泛的循环潜力，但它们会产生。 C7 的水平相对较低，并且在皮肤或粘膜中的停留情况尚未明确，因此，这是必要的。开发能够接触皮肤和粘膜皮肤组织的更有效的细胞疗法。皮肤和粘膜内丰富，并且由于其 MHC 不受限制的性质，与同种异体相容转移，但它们不会自然产生 C7。我们的创新方法将采用精密基因组。使用 CRISPR/Cas9 改造 γδ T 细胞以产生高水平的内源性 C7。 γδ T 细胞的组织迁移特性（尤其是皮肤和粘膜）以及它们表现出的同种异体相容性，使它们特别适合 C7 蛋白的治疗递送。我们将定义一种基因组工程策略，以赋予 C7 高表达、增强皮肤归巢并减少在目标 2 中，我们将评估工程化人类 γδ T 细胞的炎症/细胞毒性能力。同种异体 γδ T 细胞定位于皮肤和粘膜，沉积 C7，并改善免疫缺陷患者的病理学此外，我们将测试唑来膦酸诱导的工程体内扩增的效果。 Vγ9Vδ2 T 细胞亚群对治疗功效的影响我们的方法是一种高度新颖和创新的同种异体策略。旨在解决当前 RDEB 细胞疗法的关键局限性。 γδ T 细胞代表了一种新颖的工程和蛋白质递送策略，具有 RDEB 的转化潜力，其他遗传性皮肤粘膜疾病，以及细胞/干细胞治疗的多种不同疾病移植。

项目成果

期刊论文数量（3）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Generation and characterization of an immunodeficient mouse model of mucopolysaccharidosis type II.

II 型粘多糖贮积症免疫缺陷小鼠模型的生成和表征。

DOI：
发表时间：
2023-04
期刊：
影响因子：
3.8
作者：
Smith, Miles C;Belur, Lalitha R;Karlen, Andrea D;Podetz;Erlanson, Olivia;Laoharawee, Kanut;Furcich, Justin;Lund, Troy C;You, Yun;Seelig, Davis;Webber, Beau R;McIvor, R Scott
通讯作者：
McIvor, R Scott

A Pan-RNase Inhibitor Enabling CRISPR-mRNA Platforms for Engineering of Primary Human Monocytes.

一种泛 RNA 酶抑制剂，支持 CRISPR-mRNA 平台用于原代人单核细胞工程。

DOI：
发表时间：
2022-08-28
期刊：
影响因子：
5.6
作者：
Laoharawee, Kanut;Johnson, Matthew J;Lahr, Walker S;Sipe, Christopher J;Kleinboehl, Evan;Peterson, Joseph J;Lonetree, Cara;Bell, Jason B;Slipek, Nicholas J;Crane, Andrew T;Webber, Beau R;Moriarity, Branden S
通讯作者：
Moriarity, Branden S

Internal checkpoint regulates T cell neoantigen reactivity and susceptibility to PD1 blockade.

内部检查点调节 T 细胞新抗原反应性和对 PD1 阻断的敏感性。

DOI：
10.1016/j.medj.2022.07.008
发表时间：
2022-10-14
期刊：
Med
影响因子：
17
作者：
Palmer, Douglas C.;Webber, Beau R.;Patel, Yogin;Johnson, Matthew J.;Kariya, Christine M.;Lahr, Walker S.;Parkhurst, Maria R.;Gartner, Jared J.;Prickett, Todd D.;Lowery, Frank J.;Kishton, Rigel J.;Gurusamy, Devikala;Franco, Zulmarie;Vodnala, Suman K.;Diers, Miechaleen D.;Wolf, Natalie K.;Slipek, Nicholas J.;McKenna, David H.;Sumstad, Darin;Viney, Lydia;Henley, Tom;Burckstummer, Tilmann;Baker, Oliver;Hu, Ying;Yan, Chunhua;Meerzaman, Daoud;Padhan, Kartik;Lo, Winnie;Malekzadeh, Parisa;Jia, Li;Deniger, Drew C.;Patel, Shashank J.;Robbins, Paul F.;McIvor, R. Scott;Choudhry, Modassir;Rosenberg, Steven A.;Moriarity, Branden S.;Restifo, Nicholas P.
通讯作者：
Restifo, Nicholas P.