Developing Novel Therapies for High Risk Pediatric Cancers

Division Of Basic Sciences - Nci

Investigators

Linked publications & trials

Abstract

The overarching goal of this project is to identify, validate, and translate transformative therapeutic strategies for high-risk pediatric cancers, with a particular focus on neuroblastoma (NB). Pediatric solid tumors remain among the most treatment-resistant cancers when relapsed or refractory, necessitating the development of novel approaches that can overcome challenges such as tumor heterogeneity, immune evasion, and toxicities associated with conventional therapies. Our integrated discovery platform employs a suite of high-throughput assays-spanning genomics, epigenomics, and functional genomics-to uncover vulnerabilities and actionable targets in pediatric solid tumors. These include genome-wide CRISPR/Cas9 knockout and epigenetic perturbation screens, whole-genome and transcriptome sequencing, and dynamic live-cell monitoring platforms (e.g., Incucyte, iCELLigence). Our pharmacologic screening pipeline leverages the NCATS Mechanism Interrogation Plate, containing over 2000 annotated compounds, to systematically evaluate single and combination drug responses. Hits from these screens are evaluated in robust patient-derived xenograft (PDX) models that faithfully recapitulate human disease. In parallel, we have focused on immunotherapy-based strategies targeting pediatric solid tumors, particularly NB. Despite the success of CAR T cell therapy in hematologic malignancies, applications to solid tumors have been hindered by antigen heterogeneity, poor T cell infiltration, and immunosuppressive tumor microenvironments. Through a combination of computational target prediction, functional CAR screening, and proteomic analysis, we identified glypican-2 (GPC2) and CD276 (B7-H3) as leading targets for NB. These antigens are overexpressed on the majority of NB tumors while showing limited expression in healthy tissues. We engineered and tested a series of single-target CARs and bicistronic CARs (BiCisCARs) directed at GPC2 and CD276. CAR constructs were optimized using competitive expansion assays, CITE-seq, and in vivo xenograft modeling. BiCisCARs co-targeting GPC2 and CD276 demonstrated superior tumor clearance, prolonged persistence, and reduced T cell exhaustion compared to their monospecific counterparts. Our lead GPC2-targeting construct, CT3, has advanced to IND-enabling studies. BiCisCARs incorporating MGA271-derived CD276 binders have shown minimal off-tumor reactivity due to density-dependent killing mechanisms and are primed for future clinical development. To expand our immunotherapeutic repertoire beyond NB, we are developing dual-target CARs and novel T cell receptors (TCRs) against pediatric brain tumors and bone malignancies. These include PRAME-directed TCRs engineered with dominant-negative TGFBR2 to resist immunosuppressive signaling, currently under evaluation in preclinical models of Ewing sarcoma and osteosarcoma. Our strategy also addresses one of the central limitations of solid tumor CAR therapies-immune evasion and suppressive microenvironments. By integrating CAR T cells with polypharmacy approaches that target inhibitory myeloid components, we are enhancing the functional activity of adoptively transferred lymphocytes. These interventions are designed to remodel the tumor stroma and improve T cell infiltration and persistence. Overall, our multidisciplinary approach enables rational CAR design through data-driven selection, functional validation, and molecular optimization. Our CAR platforms exhibit hallmarks of effective cellular immunotherapy: central memory phenotype, reduced exhaustion, robust expansion, and durable tumor control. We are pioneering a new wave of adoptive cell therapies tailored to the complex immunobiology of pediatric solid tumors. In summary, this project leverages cutting-edge technologies across synthetic biology, functional genomics, and translational immunology to build and refine precision therapeutics for neuroblastoma and related pediatric malignancies. Our aim is not only to create highly specific and effective treatments but also to set a foundation for personalized pediatric oncology that transcends current therapeutic limitations.