ALK is a very promising therapeutic target in neuroblastoma, and is the focus of this application. The outcome of the research proposed here will be to generate and test a panel of ALK antibodies that are likely to be useful in the treatment of neuroblastoma, either alone or in combination with small-molecule ALK inhibitors like crizotinib. The use of monoclonal antibodies such as Herceptin/Trastuzumab or Erbitux/cetuximab to targeting oncogenic receptor tyrosine kinases such as EGFR and HER2/Neu in patients with other cancers -- alone or in combination with small molecule kinase inhibitors -- has led to significant increases in disease-free survival. However, despite the fact that ALK was identified as an oncogene in neuroblastoma 3 years ago, and is now known to be expressed on the cell surface of the majority of tumors (but not normal cells), clinically relevant antibodies for this target have not yet been developed. Our goal is to define an approach for successfully treating neuroblastoma, a deadly childhood cancer that parallels the use of Herceptin and Erbitux in breast cancer, non-small-cell lung cancer, and colorectal cancer.

We combine the expertise of a pediatric oncologist who is a specialist in understanding hereditary predisposition and progression of neuroblastoma, and who first made the discovery that ALK is mutated in a subset of patients with neuroblastoma (Dr. Mossé), with a structural biologist/biochemist who is an expert in understanding mechanisms of molecules like the ALK gene product (Dr. Lemmon). Dr. Lemmon will generate purified monoclonal antibodies that specifically target the ALK protein, and Drs. Lemmon and Mossé will together analyze which of these antibodies is most effective at killing neuroblastoma cells in culture. This collaboration will continue with studies to determine how these antibodies work, and whether they can be combined with small molecule inhibitors like crizotinib to overcome the major drug resistance ALK mutations that we have seen in neuroblastoma. Dr. Mossé's laboratory will then perform studies with the most promising antibodies in mouse models of neuroblastoma, testing predictions of the collaborative in vitro studies. The in vivo element of this project is crucial, and success will provide the preclinical rationale for development of humanized ALK antibodies for rapid clinical testing and development. With this combination of approaches, we will establish the importance of ALK-targeted antibodies as an immunotherapeutic approach for patients with highly malignant neuroblastoma and will be ready to expedite the path to a Phase 1 clinical trial by the end of this project.

Neuroblastoma remains a devastating clinical problem and continues to be a leading cause of childhood cancer morbidity and mortality despite dramatic increases in chemoradiotherapy. Defining how to attack neuroblastoma effectively, in light of the key role that ALK plays in this disease, is the primary goal of this proposal. Through the research proposed here, we will determine how to apply immunotherapeutic targeting of ALK to patients neuroblastoma -- leading to what we hope will be a dramatic change in the standard of care for these patients.