Nanobody based imaging and targeting of ECM in disease and development
Inventors
Hynes, Richard O. • Jailkhani, Noor • Ploegh, Hidde L. • Xie, Yushu J.
Assignees
Boston Childrens Hospital • Massachusetts Institute of Technology • Whitehead Institute for Biomedical Research
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Publication Number
US-12454576-B2
Publication Date
2025-10-28
Expiration Date
Abstract
Methods for developing disease-related nanobodies and related products and kits are provided. The disease-specific proteins are extracellular matrix (ECM) proteins, domains or epitopes that are associated with various aspects of disease and are not present, or are present in very low quantities, in non-diseased individuals. Highly effective nanobodies capable of specifically binding to these ECM protein epitopes useful in in vivo imaging assays, the detection, diagnosis and treatment of diseases as well as monitoring therapeutic progress in a patient with a disease are provided herein.
Core Innovation
The invention relates to disease-enriched extracellular matrix (ECM) targeting nanobodies (VHH) that bind disease state ECM epitopes present in greater amounts in diseased tissue than in normal tissue, with nM to sub-pM affinity. Exemplary disease state ECM epitopes include fibronectin EIIIA/EIIIB and tenascin C, including targeting of specific fibronectin and tenascin C domains.
The invention further relates to chimeric antigen receptor (CAR) constructs in which the ectodomain comprises a nanobody specific for the diseased state ECM epitope, along with a transmembrane domain and an endodomain. A related embodiment includes chimeric antigen receptor T cells (CAR T cells) containing the claimed CAR constructs, including VHH-based CAR-T targeting EIIIB.
In addition to CAR constructs, the document describes conjugated ECM-targeting nanobodies for imaging and diagnosis, and for detecting ECM epitopes and monitoring disease progression or regression. Imaging is described using Immuno-PET/CT with Cu-64-labeled nanobodies and visualization of primary tumors and small metastases.
Claims Coverage
The independent claims cover CAR constructs whose ectodomains bind directly to diseased state ECM epitopes present at higher amounts in diseased tissue than in normal tissue, with defined ectodomain sequence/CDR configurations plus transmembrane and endodomain elements. Two independent claim sets are present.
CAR construct with nanobody-defined CDR sets for diseased-state ECM epitopes
A CAR construct comprising an ectodomain comprising a nanobody that is specific for and binds directly to a diseased state extracellular matrix (ECM) epitope present in greater amounts in diseased tissue than in normal tissue; a transmembrane domain; and an endodomain; wherein the nanobody comprises CDR1, CDR2 and CDR3 comprising specified SEQ ID NO combinations.
CAR construct with ectodomain sequence set for diseased-state ECM epitopes
A CAR construct comprising an ectodomain comprising a sequence set forth in SEQ ID NOs: 1-4 (or an antigen binding fragment thereof) that is specific for and binds directly to a diseased state extracellular matrix (ECM) epitope present in greater amounts in diseased tissue than in normal tissue; a transmembrane domain; and an endodomain.
Across the independent claims, the inventive coverage centers on CAR constructs that directly recognize diseased-state ECM epitopes enriched in diseased tissue, where ectodomains are specified either by explicit CDR1/CDR2/CDR3 SEQ ID NO combinations or by a defined SEQ ID NO sequence set (or antigen binding fragment). Dependent claims further add binding affinity constraints and example ECM epitope targets such as the EIIIB domain of fibronectin and human Tenascin C.
Stated Advantages
Suppresses tumor growth and alters tumor vasculature and immune infiltration.
Detecting ECM epitopes and monitoring disease progression or regression.
Visualization of primary tumors and small metastases.
Documented Applications
Imaging and diagnosis.
Detecting ECM epitopes and monitoring disease progression or regression.
Immuno-PET/CT with Cu-64-labeled nanobodies.
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