Effect of fabrication parameters on morphology and drug loading of polymer particles for rosiglitazone delivery.

For the past several decades, drug-encapsulated polymer particles have been investigated as locally-delivered, long-acting therapies. The most common method of producing such particles is the oil in water solvent extraction technique. Using this technique, we produced poly(lactide-co-glycolide) (PLG) microparticles encapsulating rosiglitazone, a small molecule anti-diabetic drug. We investigated the impact of modulating fabrication parameters, including choice of organic solvent, concentration of polymer, and speed of homogenization and centrifugation on particle morphology and drug loading. Additionally, we studied the ratio of air-water-interface area to the extraction bath volume, a previously unstudied fabrication parameter, and its impact on rosiglitazone loading when using dichloromethane as the organic solvent. Under the conditions tested, drug loading can be increased 5-fold by increasing this ratio, which may be achieved by simply selecting a larger extraction vessel. By changing the organic solvent from dichloromethane to ethyl acetate, we produced particles with 60% higher rosiglitazone loading. Interestingly, the particles made with ethyl acetate appeared phase dark under light microscopy suggesting the presence of internal pores. By increasing the proportion of organic phase in the emulsion we eliminated the aberrant morphology but did not alter drug loading. As a final step in the development of the particles, we established that rosiglitazone remained stable throughout the encapsulation process and its subsequent release from particles by demonstrating that rosiglitazone loaded particles enhanced adipocyte lipid storage and adiponectin secretion. Taken together, for this system, air-water-interface area to volume ratio of the extraction bath and organic solvent both arose as key parameters in maximizing rosiglitazone loading in PLG microparticles. This study of how fabrication parameters impact drug loading and particle morphology may be useful in other investigations to encapsulate small molecules in polymer particles for controlled release applications.

Advances in immunotherapeutic targets for childhood cancers: A focus on glypican-2 and B7-H3.

Cancer immunotherapies have revolutionized how we can treat adult malignancies and are being translated to pediatric oncology. Chimeric antigen receptor T-cell therapy and bispecific antibodies targeting CD19 have shown success for the treatment of pediatric patients with B-cell acute lymphoblastic leukemia. Anti-GD2 monoclonal antibody has demonstrated efficacy in neuroblastoma. In this review, we summarize the immunotherapeutic agents that have been approved for treating childhood cancers and provide an updated review of molecules expressed by pediatric cancers that are under study or are emerging candidates for future immunotherapies. Advances in our knowledge of tumor immunology and in genome profiling of cancers has led to the identification of new tumor-specific/associated antigens. While cell surface antigens are normally targeted in a major histocompatibility complex (MHC)-independent manner using antibody-based therapies, intracellular antigens are normally targeted with MHC-dependent T cell therapies. Glypican 2 (GPC2) and B7-H3 (CD276) are two cell surface antigens that are expressed by a variety of pediatric tumors such as neuroblastoma and potentially can have a positive impact on the treatment of pediatric cancers in the clinic.

CAR T cells targeting tumor-associated exons of glypican 2 regress neuroblastoma in mice.

Targeting solid tumors must overcome several major obstacles, in particular, the identification of elusive tumor-specific antigens. Here, we devise a strategy to help identify tumor-specific epitopes. Glypican 2 (GPC2) is overexpressed in neuroblastoma. Using RNA sequencing (RNA-seq) analysis, we show that exon 3 and exons 7-10 of GPC2 are expressed in cancer but are minimally expressed in normal tissues. Accordingly, we discover a monoclonal antibody (CT3) that binds exons 3 and 10 and visualize the complex structure of CT3 and GPC2 by electron microscopy. The potential of this approach is exemplified by designing CT3-derived chimeric antigen receptor (CAR) T cells that regress neuroblastoma in mice. Genomic sequencing of T cells recovered from mice reveals the CAR integration sites that may contribute to CAR T cell proliferation and persistence. These studies demonstrate how RNA-seq data can be exploited to help identify tumor-associated exons that can be targeted by CAR T cell therapies.

The Role of Glypicans in Cancer Progression and Therapy.

Glypicans are a family of heparan sulfate proteoglycans that are attached to the cell membrane via a glycosylphosphatidylinositol anchor. Glypicans interact with multiple ligands, including morphogens, growth factors, chemokines, ligands, receptors, and components of the extracellular matrix through their heparan sulfate chains and core protein. Therefore, glypicans can function as coreceptors to regulate cell proliferation, cell motility, and morphogenesis. In addition, some glypicans are abnormally expressed in cancers, possibly involved in tumorigenesis, and have the potential to be cancer-specific biomarkers. Here, we provide a brief review focusing on the expression of glypicans in various cancers and their potential to be targets for cancer therapy.