Biology subjects

Fields, J.

Publications and source records attributed to Fields, J..

3 recordsLinked to original sources

bioRxiv2026

Mapping breast cancer lineage in radiation and immunotherapy using the REMAP mouse

We present REMAP (Recording Evolution in Mammary tumors via Active PyMT), a lineage-tracing mouse model that integrates inducible CRISPR recording with the MMTV-PyMT model of hormone receptor-positive (HR+) breast cancer. Inducible Cas9 drives editing of MARC1 homing guide RNAs (hgRNAs), generating heritable lineage marks, and enables reconstruction of clonal relationships. Using REMAP, we profiled tumor evolution and response to radiation combined with anti-PD1 immunotherapy. Treatment reduced tumor burden locally and systemically, and single-cell RNA sequencing revealed remodeling of the tumor microenvironment (TME). We identified metastatic clones present across primary tumors and distant sites, which exhibited elevated epithelial-mesenchymal transition (EMT) programs as a heritable clonal state. Treatment reduced EMT-associated transcriptional programs and reshaped immune composition, with radiation driving clonal expansion of T cells and reduced repertoire diversity. In contrast, cancer-associated fibroblast clones spanned multiple transcriptional states, indicating substantial stromal plasticity. Together, REMAP enables high-resolution coupling of clonal history and cellular state in vivo, revealing that tumor progression, metastasis, and therapeutic response are governed by heritable lineage programs.

cancer biology↗

bioRxiv2025

Effects and mechanisms of monoclonal and polyclonal human antibodies in protection of humanized mice from HIV-1 challenge

Recent clinical trials of both active and passive immunization demonstrate the barriers to the successful development of efficacious preventative HIV-1 vaccines and prophylactic antibody treatments. More facile means to explore these interventions in preclinical models could fill key gaps in knowledge and contribute to identifying and optimizing the most promising interventions. Here we report simple adaptations to standard virus challenge strategies in the humanized mouse model that support evaluation of antibody-mediated protection from infection with fewer high stakes design choices and demonstrate evaluation of protection afforded by polyclonal human serum IgG antibodies (pAbs) and monoclonal antibodies (mAbs) with variable pharmacokinetic (PK) and functional profiles. Using these adaptations, we observed that both neutralization and Fc-mediated effector functions contribute to the in vivo antiviral activity of broadly-neutralizing antibody VRC01, that confounding of results due to differences in mAb PK can be overcome, and most promisingly, that polyclonal human serum IgG that exhibits potent neutralizing and Fc-effector function can protect from infection. Collectively, this work demonstrates insights into antibody-mediated protection and methods that hold promise in supporting testing the protection from HIV-1 afforded by human pAb responses induced by vaccination.

immunology↗

bioRxiv2025

Intratumoral expression of IL-12 and CD40 ligand (CD154) from plasmids generates antitumor responses that eliminate tumoral T regs

Intratumoral immunotherapy (ITIT) strives to generate effective antitumor immunity by directly stimulating the immune system in tumors to reverse local tumor-mediated immune suppression. In vivo expression of Interleukin-12 (IL-12) using in vivo plasmid transfection as an intratumoral cancer immunotherapy entered Phase II clinical trials for metastatic melanoma but to limited clinical success. We sought to improve the efficacy of in vivo IL-12 electroporation by the addition of a CD154 (CD40 ligand)- expressing plasmid to the IL-12 encoding plasmid treatment and assessing efficacy against solid tumors. Mice with intradermal B16F10 melanoma or MC38 murine colon carcinoma tumors received 2 weekly intratumoral (IT) injections of plasmids encoding IL-12 and CD154, followed by in vivo electroporation. The addition of CD154 to IL-12 was superior to IL-12 alone and resulted in frequent tumor clearance of treated tumors, marked by an increase in CD8 T cells and a drastic reduction in T regulatory cells in the tumor microenvironment. Tumor treatment responses were abrogated in mice which lack conventional DC1 cells (BatF3 KO) or lack CD8 T cells. These findings highlight the potential of adding CD154 to IL-12 plasmid electroporation as a cancer immunotherapy and suggest that other combinations would be therapeutically valuable.

immunology↗