Single-chain antibody countermeasures for the Radiation GI Syndrome

放射胃肠道综合症的单链抗体对策

基本信息

批准号：
8852053
负责人：
Jim Rotolo
金额：
$ 10.1万
依托单位：
CERAMIDE THERAPEUTICS, LLC
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-06-01 至 2017-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8852053
关键词：
Acute Affinity Animal Model Animals Antibodies Apoptosis Atherosclerosis Attenuated Autoimmune encephalitis Binding Biological Assay Biological Availability Bioshield Biotechnology Blood Circulation Blood Vessels Bone Marrow Cell Death Cell Survival Ceramide Signaling Pathway Ceramides Cessation of life Chimeric Proteins Cloning Consensus Sequence Cytoprotection Cytotoxic T-Lymphocytes Data Development Diabetes Mellitus Digestion Directed Molecular Evolution Disasters Dose Drug Kinetics Enzyme-Linked Immunosorbent Assay Epithelial Exposure to Fab Immunoglobulins Gastrointestinal tract structure Genetic Half-Life Health Hour Human Image Immunofluorescence Immunologic Immunoglobulin Fragments Immunoglobulin G Immunotherapeutic agent In Vitro Inhibition of Apoptosis Intestines Jurkat Cells Lead Length Light Mediating Medical Membrane Modeling Monoclonal Antibodies Mus Nuclear Parents Pathology Penetration Proteins Publishing Radiation Radiation Protection Radiation Syndromes Radiation Tolerance Radiation Toxicity Radioprotection Reagent Recombinants Role Route Schedule Serum Stem cells Syndrome Techniques Testing Therapeutic Therapeutic antibodies Time Tissues Toxic effect Vascular Endothelium Whole-Body Irradiation antibody inhibitor antigen binding base complementarity-determining region 3 cost gastrointestinal graft vs host disease humanized monoclonal antibodies improved in vivo irradiation novel pre-clinical product development prophylactic research study small molecule

项目摘要

DESCRIPTION (provided by applicant): Genetic or pharmacologic inhibition of ceramide-mediated cell death within the vascular endothelium of the gastrointestinal tract (GI) protects mice from death from the Radiation GI Syndrome (RGS). Prophylactic administration of anti-ceramide antibody attenuates endothelial apoptosis, resulting in significant protection of intestinal stem cells and animal survival following lethal radiation exposure. Administration 24 h post radiation exposure similarly protects crypt stem cells and dramatically improves survival, indicating that anti-ceramide antibody represents the first effective antibody of small molecule mitigator of lethal RGS. To improve upon the efficacy of anti-ceramide antibody, we propose to select a recombinant single-chain antibody fusion protein containing the known ceramide-binding regions of anti-ceramide antibody as a novel Medical Radiation Countermeasure. Single-chain antibody fusion proteins are smaller derivatives of full length antibodies, and thus offer the advantage of rapid entering into the bloodstream and increased penetration into tissue compared to full-length antibodies. Additionally, single-chain antibody fusion proteins can be modified by general cloning techniques to display preferred target binding, and from a product development standpoint these fusion proteins can be produced easily and at minimal cost. While in some cases single-chain antibody fusion proteins may not retain full activity of the parent antibody, our preliminary data indicates that similar antibody fragments derived by enzymatic digestion of humanized anti-ceramide antibody results in significant protection and mitigation of radiation-induced crypt stem cell lethality, even 24 h after high single-dose exposure. These data indicate that a single-chain antibody fusion protein can be sufficient to induce therapeutic neutralization of ceramide. As such, a neutralizing anti-ceramide single-chain antibody fusion protein represents a promising candidate to fulfill the Project BioShield mandate for development of countermeasures to mitigate acute radiation syndromes within the first 24 h after a nuclear disaster.

描述（由申请人提供）：对胃肠道（GI）血管内皮内神经酰胺介导的细胞死亡进行遗传或药理学抑制可保护小鼠免于死于辐射胃肠道综合症（RGS）。预防性施用抗神经酰胺抗体可减弱内皮细胞凋亡，从而显着保护肠道干细胞和致命辐射暴露后的动物存活。辐射暴露后 24 小时给药同样可以保护隐窝干细胞并显着提高存活率，表明抗神经酰胺抗体代表了致死性 RGS 的小分子缓解剂的第一种有效抗体。为了提高抗神经酰胺抗体的功效，我们建议选择含有已知抗神经酰胺抗体神经酰胺结合区域的重组单链抗体融合蛋白作为新型医学辐射对策。单链抗体融合蛋白是全长抗体的较小衍生物，因此与全长抗体相比具有快速进入血流并增加对组织的渗透的优点。此外，单链抗体融合蛋白可以通过通用克隆技术进行修饰，以显示优选的靶标结合，并且从产品开发的角度来看，这些融合蛋白可以以最低的成本轻松生产。虽然在某些情况下，单链抗体融合蛋白可能无法保留亲本抗体的全部活性，但我们的初步数据表明，通过酶消化人源化抗神经酰胺抗体衍生的类似抗体片段可显着保护和减轻辐射诱导的隐窝干细胞致死率，即使在高单剂量暴露后 24 小时。这些数据表明单链抗体融合蛋白足以诱导神经酰胺的治疗性中和。因此，中和性抗神经酰胺单链抗体融合蛋白是履行 BioShield 项目任务的有前途的候选者，该项目旨在制定对策，以在核灾难后的最初 24 小时内减轻急性辐射综合症。