Fc-mediated immune stimulating, pro-inflammatory and antitumor effects of anti-HER2 IgE against HER2-expressing and trastuzumab-resistant tumors.

Abstract

Background: Anti-human epidermal growth factor receptor 2 (HER2) IgG1-based antibody therapies significantly improve cancer prognosis, yet intrinsic or acquired resistance to fragment antigen-binding (Fab)-mediated direct effects commonly occurs. Most resistant tumors retain antigen expression and therefore remain potentially targetable with anti-HER2 therapies that promote immune-mediated responses. Tumor-antigen-specific IgE class antibodies can mediate powerful immune cell-mediated effects against different cancers and have been shown to activate IgE Fc receptor-expressing monocytes. We previously reported the engineering of a trastuzumab-equivalent anti-HER2 IgE antibody and showed early evidence of Fc-mediated cancer cell-targeting effects. In the present study, we evaluated the anti-tumoral functions of two anti-HER2 IgEs, trastuzumab and pertuzumab IgE.

Methods: In vitro functionality of the two anti-HER2 antibodies was assessed by HER2 phosphorylation and ligand-independent viability assays, as well as basophil (RBL-SX38) degranulation, antibody-dependent cellular cytotoxicity/antibody-dependent cellular phagocytosis(ADCC/ADCP) assays and primary monocyte stimulation assays. The potential to trigger a hypersensitivity type I reaction was investigated using the basophil activation test (BAT). anti-tumoral efficacy was assessed in two humanized HER2+, trastuzumab-resistant models in vivo. Changes in the tumor microenvironment were assessed by flow cytometry or bulk RNA sequencing.

Results: We demonstrate the anti-tumoral and immunostimulatory functions of two anti-HER2 IgEs derived from variable region sequences of the clinically available trastuzumab and pertuzumab IgG1 antibodies. IgE engagement of monocytes via the Fc region induced tumor cell cytotoxicity and a pro-inflammatory shift with upregulation of immune-stimulatory CD40, CD80 and CD86, and downregulation of scavenger CD163, cell surface molecules. This was accompanied by enhanced pro-inflammatory tumor necrosis factor (TNF)-α, interleukin (IL)-6, IL-1β cytokine production. The absence of basophil activation by anti-HER2 IgEs ex vivo in whole blood points to potentially safe administration in humans. In two trastuzumab-resistant HER2+ tumor xenograft models in immunodeficient mice reconstituted with human immune cells, the trastuzumab-equivalent anti-HER2 IgE restricted tumor growth. Treatment was associated with enriched classical (CD14⁺CD16^-) monocyte and lower alternatively-activated (CD163⁺CD206⁺) macrophage infiltration, and higher densities of activated CD4⁺ (CD127^loCD25^hi) T cells and favorable effector T cell(Teff) to regulatory T cell (Treg) ratios in tumors.

Conclusion: Collectively, anti-HER2 IgE maintains Fab-mediated antitumor activity, induces Fc-mediated effects against HER2-expressing tumor cells, and stimulates remodeling of the immune microenvironment in tumors to promote pro-inflammatory cell phenotypes which could translate to improved outcomes for patients.