Dishevelled 2 regulates cancer cell proliferation and T cell mediated immunity in HER2-positive breast cancer

Abstract

Background Dishevelled paralogs (DVL1, 2, 3) are key mediators of Wnt pathway playing a role in constitutive oncogenic signaling influencing the tumor microenvironment. While previous studies showed correlation of β-catenin with T cell gene expression, little is known about the role of DVL2 in modulating tumor immunity. This study aimed to uncover the novel interaction between DVL2 and HER2-positive (HER2+) breast cancer (BC) in regulating tumor immunity and disease progression. Methods DVL2 loss of function studies were performed with or without a clinically approved HER2 inhibitor, Neratinib in two different HER2+ BC cell lines. We analyzed RNA (RT-qPCR) and protein (western blot) expression of classic Wnt markers and performed cell proliferation and cell cycle analyses by live cell imaging and flow cytometry, respectively. A pilot study in 24 HER2+ BC patients was performed to dissect the role of DVL2 in tumor immunity. Retrospective chart review on patient records and banked tissue histology were performed. Data were analyzed in SPSS (version 25) and GraphPad Prism (version 7) at a significance p < 0.05. Results DVL2 regulates the transcription of immune modulatory genes involved in antigen presentation and T cell maintenance. DVL2 loss of function down regulated mRNA expression of Wnt target genes involved in cell proliferation, migration, invasion in HER2+ BC cell lines (±Neratinib). Similarly, live cell proliferation and cell cycle analyses reveal that DVL2 knockdown (±Neratinib) resulted in reduced proliferation, higher growth arrest (G1), limited mitosis (G2/M) compared to non-targeted control in one of the two cell lines used. Analyses on patient tissues who received neoadjuvant chemotherapy (n = 14) further demonstrate that higher DVL2 expression at baseline biopsy pose a significant negative correlation with % CD8α levels (r = − 0.67, p < 0.05) while have a positive correlation with NLR (r = 0.58, p < 0.05), where high NLR denotes worse cancer prognosis. These results from our pilot study reveal interesting roles of DVL2 proteins in regulating tumor immune microenvironment and clinical predictors of survival in HER2+ BC. Conclusion Our study demonstrates potential immune regulatory role of DVL2 proteins in HER2+ BC. More in-depth mechanistic studies of DVL paralogs and their influence on anti-tumor immunity may provide insight into DVLs as potential therapeutic targets benefiting BC patients.

Dishevelled localization and function are differentially regulated by structurally distinct sterols

“…Nuclear DVL proteins also exhibit Wnt-independent functions. ChIP sequencing demonstrated strong association of DVL1 and DVL3 with promoters for genes that regulate the cell cycle, chromatin organization, DNA repair, signal transduction, and immune system modulation (Boligala et al, 2022; Castro-Piedras et al, 2021; Rasha et al, 2023). Nuclear DVL1 and DVL3 were also essential for myoblast proliferation in a β-catenin-independent manner (Pruller et al, 2022).…”

“…Mutation of DVL isoforms in animal models cause defects in the development of the cardiovascular tract, neural crest, skeleton and cochlear systems (Etheridge et al, 2008; Hamblet et al, 2002) while autosomal dominant mutations in DVL isoforms lead to Robinow syndrome, characterized by broad malformations within the skeletal system, urogenital system, and neurological deficits (Schwartz et al, 2021; White et al, 2015; White et al, 2016). Lastly, changes in DVL isoform expression and localization have been reported in cancers including glioma (Li et al, 2014), colorectal (Metcalfe et al, 2010), and breast cancer (Metcalfe et al, 2010; Rasha et al, 2023).…”

Automated Detection and Scoring of Tumor-Infiltrating Lymphocytes in Breast Cancer Histopathology Slides

“…Several methods are used for breast cancer treatments including surgery, hormonal therapy, and radiation therapy [3]. One of the newly developed methods for breast cancer treatment is immunotherapy, where immune cells are extracted from the patient's body and grown in the laboratory, then injected back into the patient to detect and kill tumor cells [4]. These immune cells are called stromal tumor-infiltrating lymphocytes (sTILs).…”