Simultaneous selection of nanobodies for accessible epitopes on immune cells in the tumor microenvironment.

Autor: Sekar TV; Department of Radiation Oncology, the University of Texas Southwestern Medical Center, Dallas, TX, USA.; Department of Microbiology, Pondicherry University, Kalapet, Puducherry, India., Elghonaimy EA; Department of Radiation Oncology, the University of Texas Southwestern Medical Center, Dallas, TX, USA., Swancutt KL; Department of Radiation Oncology, the University of Texas Southwestern Medical Center, Dallas, TX, USA., Diegeler S; Department of Radiation Oncology, the University of Texas Southwestern Medical Center, Dallas, TX, USA., Gonzalez I; Department of Radiation Oncology, the University of Texas Southwestern Medical Center, Dallas, TX, USA., Hamilton C; Department of Radiation Oncology, the University of Texas Southwestern Medical Center, Dallas, TX, USA., Leung PQ; Department of Radiation Oncology, the University of Texas Southwestern Medical Center, Dallas, TX, USA., Meiler J; Department of Chemistry and Center for Structural Biology, Vanderbilt University, Nashville, TN, USA.; Institute for Drug Discovery, Leipzig University Medical School, Leipzig, Germany., Martina CE; Department of Chemistry and Center for Structural Biology, Vanderbilt University, Nashville, TN, USA., Whitney M; Department of Pharmacology, University of California San Diego, La Jolla, CA, USA., Aguilera TA; Department of Radiation Oncology, the University of Texas Southwestern Medical Center, Dallas, TX, USA. todd.aguilera@utsouthwestern.edu.
Jazyk: angličtina
Zdroj: Nature communications [Nat Commun] 2023 Nov 17; Vol. 14 (1), pp. 7473. Date of Electronic Publication: 2023 Nov 17.
DOI: 10.1038/s41467-023-43038-z
Abstrakt: In the rapidly advancing field of synthetic biology, there exists a critical need for technology to discover targeting moieties for therapeutic biologics. Here we present INSPIRE-seq, an approach that utilizes a nanobody library and next-generation sequencing to identify nanobodies selected for complex environments. INSPIRE-seq enables the parallel enrichment of immune cell-binding nanobodies that penetrate the tumor microenvironment. Clone enrichment and specificity vary across immune cell subtypes in the tumor, lymph node, and spleen. INSPIRE-seq identifies a dendritic cell binding clone that binds PHB2. Single-cell RNA sequencing reveals a connection with cDC1s, and immunofluorescence confirms nanobody-PHB2 colocalization along cell membranes. Structural modeling and docking studies assist binding predictions and will guide nanobody selection. In this work, we demonstrate that INSPIRE-seq offers an unbiased approach to examine complex microenvironments and assist in the development of nanobodies, which could serve as active drugs, modified to become drugs, or used as targeting moieties.
(© 2023. The Author(s).)
Databáze: MEDLINE
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