Generation of Therapeutic Human Anti-Trop-2 Extracellular Domain Antibodies (UCLA Case No. 2026-026)

UCLA researchers from the Department of Molecular and Medical Pharmacology and the Department of Microbiology, Immunology and Molecular Genetics have developed a panel of human antibodies and chimeric antigen receptors targeting Trop-2, providing diagnostic and therapeutic resources for targeting Trop2-driven cancer.

BACKGROUND: Trophoblast cell surface antigen 2 (Trop-2) is a type-I transmembrane protein that has emerged as a clinically relevant oncogenic driver in nearly all epithelial cancers. Prior studies have demonstrated that Trop2 undergoes regulated intramembrane proteolysis, releasing its extracellular domain and producing a nuclear-localized intracellular domain that promotes tumor initiation and self-renewal. Although several pharmaceutical companies have developed anti-Trop-2 antibodies with preclinical efficacy, these constructs are typically murine or chimeric, which increases the likelihood of immunogenicity and limits their long-term therapeutic utility. Currently, two FDA-approved Trop2-targeting ADCs are available for breast cancer: sacituzumab govitecan (SG/Trodelvy®) and datopotamab deruxtecan (Dato-DXd/Datroway®)7,8. However, they have modest efficacy or concerning safety profiles and both have TOP1 inhibitors as payloads, and emergence of TOP1 mutations can confer therapeutic resistance. Thus, there is an unmet need for a fully human antibody platform with improved specificity, efficacy, minimal side effects and reduced immunogenicity.

INNOVATION: Dr. Tanya Stoyanova at the Department of Molecular and Medical Pharmacology and Dr. Owen Witte at the Department of Microbiology, Immunology and Molecular Genetics at UCLA, together with collaborators have generated the first fully human anti-Trop-2extracellular domain antibodies through high-throughput phage display technology. Using a human single-fold scFv library developed at UCLA, the inventors isolated 23 unique antibody clones that specifically bind the Trop2 extracellular domain with varying affinities. These antibodies were expressed as scFv-Fc fusion proteins in mammalian cells, purified, and quantitatively ranked for binding strength using FACS analysis across prostate cancer cell lines expressing different Trop2 levels. These antibodies exhibit high specificity and potent inhibition of Trop2 activation and downstream signaling. Further, researchers conjugated antibodies to indocyanine green (ICG), an FDA-approved optical imaging dye, to generate anti-Trop2 antibody fragment ICG conjugates that can provide low cost, high-sensitivity optical detection and intraoperative guidance in Trop2+ cancers. Additionally, five representative antibodies with distinct binding strengths were incorporated into chimeric antigen receptor (CAR) constructs. Two of these Trop2-CARs exhibited potent and selective cytotoxicity toward Trop2-overexpressing tumor cells while sparing those with endogenous expression levels. Collectively, this platform provides a versatile group of human antibody, CAR, and imaging modalities for Trop2 targeted therapy, diagnostics, and surgical applications.

POTENTIAL APPLICATIONS:

  • Development of Trop-2-targeted antibodies, antibody-drug conjugates, bi-specific antibodies, and CAR-T therapies
  • Diagnostic imaging and quantification of Trop-2 expression in tumors and patient samples
  • Optical tumor visualization and lymph-node mapping during surgery
  • Molecular tools for studying Trop-2 signaling, localization, and activation

ADVANTAGES:

  • Fully human antibodies minimize immunogenicity and are compatible with clinical translation
  • Validated in multiple Trop-2+ carcinoma models
  • Compatibility with therapeutic, diagnostic, and imaging modalities
  • Potential to block Trop-2 cleavage, activation, and downstream oncogenic signaling
  • Strong specificity and reproducibility in cell-based models

DEVELOPMENT-TO-DATE: UCLA researchers have developed 23 anti-human Trop-2 extracellular domain antibodies using phage display technology and shown that they can be used to detect human Trop-2 on the cell surface of cancer cells. The naked anti-human Trop-2 antibodies exhibit strong efficacy in pre-clinical models with no measurable side effects. They have also generated 5 Trop-2 chimeric antigen receptors that may be used for cancer immunotherapies, as well as Trop-2 antibody-ICG conjugates for intraoperative optical guidance.

Related Papers (from the inventors only):

  1. Tanya Stoyanova, Andrew Goldstein, Houjian Cai, Justin M. Drake, Jiaoti Huang, Hong Zhang, Owen N. Witte; Abstract C70: Trop2 regulates prostate tumorigenesis and stem cell self-renewal. Cancer Res 6 February 2012; 72 (4_Supplement): C70. https://doi.org/10.1158/1538-7445.PRCA2012-C70
  2. Sinawang PD, Ozen MO, Liu S, Hsu EC, Akin D, Ding E, Nolley R, Brooks JD, Stoyanova T, Demirci U. Extracellular Vesicles in Serum Carry Trop2 Protein as a Potential Molecular Indicator in Prostate Cancer. J Extracell Biol. 2025 Sep 23;4(9):e70083. doi: 10.1002/jex2.70083. PMID: 40994721; PMCID: PMC12456249.
  3. Liu S, Hsu EC, Aslan M, Garcia-Marques F, Shen M, Hartono AB, Solano F, Le K, Hwang H, Lee CS, Bermudez A, Nolley R, Peehl DM, Brooks JD, Liss MA, Pitteri SJ, Stoyanova T. Extracellular Domain Shedding of TROP2 Activates EGFR Signaling to Drive Prostate Cancer Metastasis. Cancer Res. 2025 Sep 5. doi: 10.1158/0008-5472.CAN-24-4855. Epub ahead of print. PMID: 40911783.
  4. Liu S, Hawley SJ, Kunder CA, Hsu EC, Shen M, Westphalen L, Auman H, Newcomb LF, Lin DW, Nelson PS, Feng Z, Tretiakova MS, True LD, Vakar-Lopez F, Carroll PR, Simko J, Gleave ME, Troyer DA, McKenney JK, Brooks JD, Liss MA, Stoyanova T. High expression of Trop2 is associated with aggressive localized prostate cancer and is a candidate urinary biomarker. Sci Rep. 2024 Jan 4;14(1):486. doi: 10.1038/s41598-023-50215-z. PMID: 38177207; PMCID: PMC10766957.
  5. Hsu EC, Rice MA, Bermudez A, Marques FJG, Aslan M, Liu S, Ghoochani A, Zhang CA, Chen YS, Zlitni A, Kumar S, Nolley R, Habte F, Shen M, Koul K, Peehl DM, Zoubeidi A, Gambhir SS, Kunder CA, Pitteri SJ, Brooks JD, Stoyanova T. Trop2 is a driver of metastatic prostate cancer with neuroendocrine phenotype via PARP1. Proc Natl Acad Sci U S A. 2020 Jan 28;117(4):2032-2042. doi: 10.1073/pnas.1905384117. Epub 2020 Jan 13. PMID: 31932422 
  6. Stoyanova T, Goldstein AS, Cai H, Drake JM, Huang J, Witte ON. Regulated proteolysis of Trop2 drives epithelial hyperplasia and stem cell self-renewal via beta-catenin signaling. Genes Dev. 2012 Oct 15;26(20):2271-85. doi: 10.1101/gad.196451.112. PMID: 23070813

TDG Keywords:

Diagnostic Markers > Cancer

Diagnostic Markers > Targets and Assays

Life Science Research Tools > Antibodies

Therapeutics > Oncology

Therapeutics > Immunology and Immunotherapy

 

Keywords: Trop-2, antibody, CAR-T, imaging, ICG, epithelial cancer, phage display, immunotherapy, prostate cancer, antibody-drug conjugate, therapeutic antibody, cancer diagnostics

Patent Information: