S C Saunderson, J C Halpin, G M Y Tan, P Shrivastava, A D McLellan
{"title":"Conversion of anti-tissue factor antibody sequences to chimeric antigen receptor and bi-specific T-cell engager format.","authors":"S C Saunderson, J C Halpin, G M Y Tan, P Shrivastava, A D McLellan","doi":"10.1007/s00262-024-03778-3","DOIUrl":null,"url":null,"abstract":"<p><strong>Background: </strong>The efficacy of antibody-targeted therapy of solid cancers is limited by the lack of consistent tumour-associated antigen expression. However, tumour-associated antigens shared with non-malignant cells may still be targeted using conditionally activated-antibodies, or by chimeric antigen receptor (CAR) T cells or CAR NK cells activated either by the tumour microenvironment or following 'unlocking' via multiple antigen-recognition. In this study, we have focused on tissue factor (TF; CD142), a type I membrane protein present on a range of solid tumours as a basis for future development of conditionally-activated BiTE or CAR T cells. TF is frequently upregulated on multiple solid tumours providing a selective advantage for growth, immune evasion and metastasis, as well as contributing to the pathology of thrombosis via the extrinsic coagulation pathway.</p><p><strong>Methods: </strong>Two well-characterised anti-TF monoclonal antibodies (mAb) were cloned into expression or transposon vectors to produce single chain (scFv) BiTE for assessment as CAR and CD28-CD3-based CAR or CD3-based BiTE. The affinities of both scFv formats for TF were determined by surface plasmon resonance. Jurkat cell line-based assays were used to confirm the activity of the BiTE or CAR constructs.</p><p><strong>Results: </strong>The anti-TF mAb hATR-5 and TF8-5G9 mAb were shown to maintain their nanomolar affinities following conversion into a single chain (scFv) format and could be utilised as CD28-CD3-based CAR or CD3-based BiTE format.</p><p><strong>Conclusion: </strong>Because of the broad expression of TF on a range of solid cancers, anti-TF antibody formats provide a useful addition for the development of conditionally activated biologics for antibody and cellular-based therapy.</p>","PeriodicalId":9595,"journal":{"name":"Cancer Immunology, Immunotherapy","volume":null,"pages":null},"PeriodicalIF":4.6000,"publicationDate":"2024-08-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11303627/pdf/","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Cancer Immunology, Immunotherapy","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.1007/s00262-024-03778-3","RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"IMMUNOLOGY","Score":null,"Total":0}
引用次数: 0
Abstract
Background: The efficacy of antibody-targeted therapy of solid cancers is limited by the lack of consistent tumour-associated antigen expression. However, tumour-associated antigens shared with non-malignant cells may still be targeted using conditionally activated-antibodies, or by chimeric antigen receptor (CAR) T cells or CAR NK cells activated either by the tumour microenvironment or following 'unlocking' via multiple antigen-recognition. In this study, we have focused on tissue factor (TF; CD142), a type I membrane protein present on a range of solid tumours as a basis for future development of conditionally-activated BiTE or CAR T cells. TF is frequently upregulated on multiple solid tumours providing a selective advantage for growth, immune evasion and metastasis, as well as contributing to the pathology of thrombosis via the extrinsic coagulation pathway.
Methods: Two well-characterised anti-TF monoclonal antibodies (mAb) were cloned into expression or transposon vectors to produce single chain (scFv) BiTE for assessment as CAR and CD28-CD3-based CAR or CD3-based BiTE. The affinities of both scFv formats for TF were determined by surface plasmon resonance. Jurkat cell line-based assays were used to confirm the activity of the BiTE or CAR constructs.
Results: The anti-TF mAb hATR-5 and TF8-5G9 mAb were shown to maintain their nanomolar affinities following conversion into a single chain (scFv) format and could be utilised as CD28-CD3-based CAR or CD3-based BiTE format.
Conclusion: Because of the broad expression of TF on a range of solid cancers, anti-TF antibody formats provide a useful addition for the development of conditionally activated biologics for antibody and cellular-based therapy.
背景:由于缺乏一致的肿瘤相关抗原表达,抗体靶向治疗实体瘤的疗效受到限制。不过,与非恶性细胞共有的肿瘤相关抗原仍可通过条件激活抗体或由肿瘤微环境激活的嵌合抗原受体(CAR)T细胞或通过多重抗原识别 "解锁 "后激活的CAR NK细胞进行靶向治疗。在这项研究中,我们重点研究了组织因子(TF;CD142),这是一种存在于多种实体瘤上的 I 型膜蛋白,是未来开发条件激活型 BiTE 或 CAR T 细胞的基础。TF在多种实体瘤上经常上调,为肿瘤的生长、免疫逃避和转移提供了选择性优势,并通过外凝血途径促成了血栓形成的病理过程:方法:将两种特性良好的抗血小板单克隆抗体(mAb)克隆到表达载体或转座子载体中,以产生单链(scFv)BiTE,用于评估CAR和基于CD28-CD3的CAR或基于CD3的BiTE。两种 scFv 格式对 TF 的亲和力都是通过表面等离子共振测定的。用基于 Jurkat 细胞系的检测来确认 BiTE 或 CAR 构建物的活性:结果:抗TF mAb hATR-5和TF8-5G9 mAb在转化为单链(scFv)形式后保持了纳摩尔级的亲和力,可用作基于CD28-CD3的CAR或基于CD3的BiTE形式:结论:由于TF在一系列实体瘤中广泛表达,抗TF抗体格式为开发条件激活生物制剂提供了有用的补充,可用于基于抗体和细胞的治疗。
期刊介绍:
Cancer Immunology, Immunotherapy has the basic aim of keeping readers informed of the latest research results in the fields of oncology and immunology. As knowledge expands, the scope of the journal has broadened to include more of the progress being made in the areas of biology concerned with biological response modifiers. This helps keep readers up to date on the latest advances in our understanding of tumor-host interactions.
The journal publishes short editorials including "position papers," general reviews, original articles, and short communications, providing a forum for the most current experimental and clinical advances in tumor immunology.