A Syngeneic ErbB2 Mammary Cancer Model for Preclinical Immunotherapy Trials

Abstract

In order to develop a practical model of breast cancer, with in vitro and syngeneic-immune intact in vivo growth capacity, we established a primary cell line derived from a mammary carcinoma in the transgenic FVB/N-Tg(MMTV-ErbB2*)NDL2-5Mul mouse, referred to as "NDL UCD ". The line was adapted to standard cell culture and can be transplanted into syngeneic FVB/N mice. The line maintained a stable phenotype over multiple in vitro passages and rounds of in vivo transplantation. NDL UCD tumors in FVB/N mice exhibited high expression of ErbB2 and ErbB3 and signaling molecules downstream of ErbB2. The syngeneic transplant tumors elicited an immune reaction in the adjacent stroma, detected and characterized using histology, immunophenotyping, and gene expression. NDL UCD cells also expressed PD-L1 in vivo and in vitro, and in vivo transplants were reactive to anti-immune checkpoint therapy with responses conducive to immunotherapy studies. This new NDL UCD cell line model is a practical alternative to the more commonly used 4T1 cells, and our previously described FVB/N-Tg(MMTV-PyVT)634Mul derived Met-1 fvb2 and FVB/NTg(MMTV-PyVT Y315F/Y322F) derived DB-7 fvb2 cell lines. The NDL UCD cells remained genetically and phenotypically stable over many generations, with consistent and reproducible results in immune intact preclinical cohorts.

Targeting the Aryl Hydrocarbon Receptor Signaling Pathway in Breast Cancer Development

“…Mammary epithelial cells (UCD-PYMT) were isolated from the mammary tumor of a 26-week-old B6.FVBTg(MMTV PyVT)634Mul/LellJ (PyMT) hemizygous mouse (Jackson Laboratory, Bar Harbor, ME, USA) ( 25 , 26 ) as described in Pénzváltó et al ( 27 ). Briefly, the mammary tumor was washed twice in PBS (Invitrogen, Carlsbad, CA, USA) before it was mechanically dissociated and minced in a solution of serum-free DMEM:F12 (Invitrogen) with HEPES (Invitrogen), supplemented with 0.5 mg/ml Penicillin/Streptomycin (Invitrogen), 2% bovine serum albumin fraction V (Invitrogen), 5 μg/ml insulin (Sigma Aldrich, Saint Louis, MO), 10 ng/ml cholera toxin (Sigma Aldrich, USA), and 3 mg/ml collagenase (Worthington Biochemical Corp., Lakewood, NJ).…”

In Vivo Modeling of Tumor Heterogeneity for Immuno‐Oncology Studies: Failures, Improvements, and Hopes

“…One murine melanoma cell line, YUMMER1.7, elicits T‐cell‐dependent tumor regression at low cell numbers, while escaping tumors are sensitive to anti‐cytotoxic T‐lymphocyte‐associated protein 4 (CTLA‐4) and anti‐PD‐1 combination ICB (Wang et al., 2017). Additionally, the human epidermal growth factor receptor 2 (HER2)‐driven NDL UCD cell line was recently generated as one of the few murine mammary tumor models that benefits from anti‐PD‐1 therapy (Pénzváltó et al., 2019; Silvestrini et al., 2017). As more syngeneic models are being developed, it is fundamental that a detailed characterization of their TME and response to therapy is conducted to maximize translational potential.…”

Mouse Tumor Models for Advanced Cancer Immunotherapy

“…Transplantation of in vitro cultured tumor cells into immunocompetent mice is the oldest and most commonly utilized approach to investigate antitumor therapy, including immunotherapy ( Figure 1A) [15]. Spontaneous, carcinogenic or transgenic tumor cell lines can be transplanted into such mouse strains as C57BL/6, BALB/c and FVB [16][17][18][19][20]. The creation of such models takes a short period of time, since transplanted subcutaneously or intravenously cells grow in the animal within a few weeks [21].…”