Novel nanobody development to boost immunotherapy
Mentor Name: Hui Feng
Increased MYCN oncoprotein activity consists of many difficult-to-treat neuroblastomas, childhood tumors with immunosuppressive tumor microenvironment (TME) and distant metastases. Our preliminary work has demonstrated CKLF to be highly-expressed across many solid pediatric tumors and predicts poor patient survival, especially for neuroblastoma. CKLF is upregulated by MYCN in tumor cells and secreted into the TME to attract CCR4-expressing Tregs. The CCR4 antagonist, but not commercial CKLF antibodies, disrupts the recruitment of Tregs by CKLF. We have found that genetically depleting Tregs or CKLF in zebrafish abolishes CKLF’s immunorestrictive effects, leading to enhanced anti-tumor immune responses and delaying tumor development. Our studies pinpoint a novel therapeutic strategy to restore anti-tumor immunity by targeting CKLF-mediated tumor-Treg crosstalk through CKLF-neutralizing nanobodies. The specific goal of this study is to develop and select nanobodies generated in camels to neutralize CKLF and disrupt tumor-TME crosstalk mediated by CCR4 . We will generate his-CKLF1 protein and submit the protein to an external contract-research-organization to generate nanobodies for CKLF. To select the assay to screen nanobodies, we will use 293T and neuroblastoma cells overexpressing CCR4 to determine how his-CKLF1 activates CCR4 signaling and identify the downstream readout proteins.
