Anti-EGFR VHH-armed death receptor ligand–engineered allogeneic stem cells have therapeutic efficacy in diverse brain metastatic breast cancers

Abstract

Basal-like breast cancer (BLBC) shows brain metastatic (BM) capability and overexpresses EGFR and death-receptors 4/5 (DR4/5); however, the anatomical location of BM prohibits efficient drug-delivery to these targetable markers. In this study, we developed BLBC-BM mouse models featuring different patterns of BMs and explored the versatility of estem cell (SC)–mediated bi-functional EGFR and DR4/5-targeted treatment in these models. Most BLBC lines demonstrated a high sensitivity to EGFR and DR4/5 bi-targeting therapeutic protein, EVDRL [anti-EGFR VHH (EV) fused to DR ligand (DRL)]. Functional analyses using inhibitors and CRISPR-Cas9 knockouts revealed that the EV domain facilitated in augmenting DR4/5-DRL binding and enhancing DRL-induced apoptosis. EVDRL secreting stem cells alleviated tumor-burden and significantly increased survival in mouse models of residual-tumor after macrometastasis resection, perivascular niche micrometastasis, and leptomeningeal metastasis. This study reports mechanism based simultaneous targeting of EGFR and DR4/5 in BLBC and defines a new treatment paradigm for treatment of BM.

Gene-edited and -engineered stem cell platform drives immunotherapy for brain metastatic melanomas

“…However, clinical benefits, such as longer survival, were limited to patients with LM derived from breast cancer. We previously established LM mouse models using patient-derived breast cancer cells and showed that SC delivery of targeted therapeutics is a promising approach for LM ( 57 ). In this study, we engineered SC N1KO secreting GM-CSF and scFvPD-1 to treat immunosuppressive PTEN-deficient melanoma LM mouse models and investigated the immune profile and therapeutic efficacy after IT injection of TSC-G/P.…”

Engineered cell-based therapies in ex vivo ready-made CellDex capsules have therapeutic efficacy in solid tumors

“…Human SCs (UC Davis) were grown in DMEM supplemented with 10% (vol/vol) FBS, 1% (vol/vol) L-glutamine, 1% (vol/vol) P/S, and recombinant human fibroblast growth factor hFGF (40 ng/mL; R&D Systems). Both RAW264.7 (ATCC) and CT2A (ATCC) murine cell lines and the human cell lines BMET0.2 [ 44 ], LN229-IL13Ra2, LN229-EGFRVIII and MDA-BM-231 (J. Massagué, Memorial Sloan Kettering Cancer Center), were cultured in DMEM supplemented with 10% (vol/vol) FBS and 1% (vol/vol) P/S. The NK92 (ATCC) cell line was cultured in alpha minimum essential medium (α-MEM), L-glutamine 1% (vol/vol), 1.5 g/L sodium bicarbonate, 0.2 mM inositol, 0.1 mM 2-mecroptoethanol, 0.1 mM folic acid, 12.5% horse serum, 12.5% (vol/vol) FBS, 1% (vol/vol) P/S and 100 U/mL hIL-2.…”

Nanobodies; new molecular instruments with special specifications for targeting, diagnosis and treatment of triple-negative breast cancer

“…The combination of nanobodies with another ligand or molecule has been used in many studies, and its benefits in the treatment of various diseases have been investigated. In a landmark study by Kitamura et al [ 23 ], a combination of anti-EGFR nanobody and TNF-related apoptosis-inducing ligand (TRAIL) that binds to the DR4/5 receptor was used to treat breast cancer. In this project, the effect of each of these factors on breast cancer cells was investigated, and it was shown that anti-EGFR nanobody and death receptor ligand (DRL) alone have little apoptotic effect on cancer cells, but when these two molecules are joined together, they can effectively induce apoptosis in TNBC cells.…”

“…These mesenchymal stem cells were used in mouse models of residual tumor after macro-metastasis resection, perivascular niche micro-metastasis, and leptomeningeal metastasis. The results showed that the secretion of the designed molecule by mesenchymal stem cells reduced the tumor volume and increased the survival of mice [ 23 ].…”