Extracellular domain, hinge, and transmembrane determinants affecting surface CD4 expression of a novel anti-HIV chimeric antigen receptor (CAR) construct.
| العنوان: | Extracellular domain, hinge, and transmembrane determinants affecting surface CD4 expression of a novel anti-HIV chimeric antigen receptor (CAR) construct. |
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| المؤلفون: | Zenere G; Tulane National Primate Research Center, Covington, LA 70433.; BioMedical Sciences Program, Tulane University School of Medicine, New Orleans, LA 70112., Wu C; Tulane National Primate Research Center, Covington, LA 70433., Midkiff CC; Tulane National Primate Research Center, Covington, LA 70433., Johnson NM; Tulane National Primate Research Center, Covington, LA 70433.; BioMedical Sciences Program, Tulane University School of Medicine, New Orleans, LA 70112., Grice CP; Tulane National Primate Research Center, Covington, LA 70433.; Department of Pharmacology, Tulane University School of Medicine, New Orleans, LA 70112., Wimley WC; Department of BioChemistry and Molecular Biology, Tulane University School of Medicine, New Orleans, LA 70112., Kaur A; Tulane National Primate Research Center, Covington, LA 70433.; Department of Microbiology and Immunology, Tulane University School of Medicine, New Orleans, LA 70112., Braun SE; Tulane National Primate Research Center, Covington, LA 70433.; Department of Pharmacology, Tulane University School of Medicine, New Orleans, LA 70112. |
| المصدر: | BioRxiv : the preprint server for biology [bioRxiv] 2023 Oct 26. Date of Electronic Publication: 2023 Oct 26. |
| نوع المنشور: | Preprint |
| اللغة: | English |
| بيانات الدورية: | Country of Publication: United States NLM ID: 101680187 Publication Model: Electronic Cited Medium: Internet ISSN: 2692-8205 (Electronic) Linking ISSN: 26928205 NLM ISO Abbreviation: bioRxiv Subsets: PubMed not MEDLINE |
| مستخلص: | Chimeric antigen receptor (CAR)-T cells have demonstrated clinical potential, but current receptors still need improvements to be successful against chronic HIV infection. In this study, we address some requirements of CAR motifs for strong surface expression of a novel anti-HIV CAR by evaluating important elements in the extracellular, hinge, and transmembrane (TM) domains. When combining a truncated CD4 extracellular domain and CD8α hinge/TM, the novel CAR did not express extracellularly but was detectable intracellularly. By shortening the CD8α hinge, CD4-CAR surface expression was partially recovered and addition of the LYC motif at the end of the CD8α TM fully recovered both intracellular and extracellular CAR expression. Mutation of LYC to TTA or TTC showed severe abrogation of CAR expression by flow cytometry and confocal microscopy. Additionally, we determined that CD4-CAR surface expression could be maximized by the removal of FQKAS motif at the junction of the extracellular domain and the hinge region. CD4-CAR surface expression also resulted in cytotoxic CAR T cell killing of HIV Env + target cells. In this study, we identified elements that are crucial for optimal CAR surface expression, highlighting the need for structural analysis studies to establish fundamental guidelines of CAR designs. Competing Interests: Competing interest The authors report no conflict of interest that may arise from this research. |
| التعليقات: | Update in: PLoS One. 2024 Aug 12;19(8):e0293990. doi: 10.1371/journal.pone.0293990. (PMID: 39133676) |
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| معلومات مُعتمدة: | F30 AI150452 United States AI NIAID NIH HHS; P51 OD011104 United States OD NIH HHS; R01 AI102693 United States AI NIAID NIH HHS; R21 AI145642 United States AI NIAID NIH HHS |
| فهرسة مساهمة: | Keywords: CAR structure; CAR-T cell therapy; CD8α; D1D2; chimeric antigen receptor; hinge; transmembrane domain |
| تواريخ الأحداث: | Date Created: 20231114 Latest Revision: 20240820 |
| رمز التحديث: | 20240820 |
| مُعرف محوري في PubMed: | PMC10634810 |
| DOI: | 10.1101/2023.10.25.563930 |
| PMID: | 37961145 |
| قاعدة البيانات: | MEDLINE |
| تدمد: | 2692-8205 |
|---|---|
| DOI: | 10.1101/2023.10.25.563930 |