Cambridge Hospital

Introduction/BackgroundThe observed clinical benefit of immunotherapy in EOC is limited. During cancer progression, tumor-specific T cells display increased expression of inhibitory receptors (IRs), causing their functional exhaustion. We hypothesize that protecting tumor-specific T cells from immunosuppression by gene editing approaches aimed at reducing IRs expression might maximize their therapeutic potential.MethodologyWe characterized by flow cytometry EOC cell lines and patients’ ascites-derived primary cultures (PC) to evaluate their expression of selected tumor-associated antigens and IR ligands. Considering antigen expression, we selected a HLA-A*02:01-restricted WT1-specific TCR from our library (HD1-TCR) and tested different T-cell products engineered to express HD1-TCR and also harboring gene disruption ( HD1-IRKO) of distinct IRs (2B4, TIM3, LAG3).To functionally characterize the putative advantage provided by IR disruption on the anti-tumor efficacy of engineered T cells, HD1-TCR and HD1-IRKO gene-edited T cells were challenged with WT-1pos HLA-A*02:01pos (or HLA-A*02:01neg, as control) cancer cell lines, PC and tumor derived organoids (PDOs). We evaluated cytokine secretion, T cell activation and apoptosis.ResultsWe observed high expression of WT-1 while IR ligands (IR-L) were expressed by a low percentage of tumor cells and increased by IFNγ exposure. After co-culture with tumor cells, the different T-cell products displayed similar killing of cell lines, while increased killing was observed with HD1-IRKO cells against EOC primary cultures expressing IR-Ls. In particular, TIM-3KO HD1-TCR edited T cells displayed more potent killing of PC expressing TIM-3 IR ligands compared to the other T cell products. Additionally, upon co-culture with HLA-matched PDOs, both TIM-3KO and LAG-3KO HD1-TCR T cells showed an increased induction of cancer cell apoptosis compared to IR-competent cells.ConclusionHD1-IRKO T-cell products appear superior to HD1-TCR IR-competent counterparts in killing WT-1pos HLA-A*02:01 cancer cells. The most consistent advantages were observed with TIM-3KO HD1-TCR.DisclosuresReceipt of grants/research supports from Intellia Therapeutics.Receipt of grants/research supports from GSK, MSD/Astrazeneca, Pharmamar

An important goal of early childhood education is teaching emotional self-regulation within the context of a safe, stable, nurturing environment. Expulsion of young children ignores underlying emotional and behavioral concerns, disproportionately affects children of color (Black or Hispanic), males, children with disabilities, and socioeconomically disadvantaged populations, and has long-term consequences on educational and life success. Addressing implicit bias and providing child mental health consultation (psychologists, social workers, developmental behavioral pediatricians, child psychiatrists, and child neurologists) to child care providers can prevent expulsion. Pediatricians and other providers within the medical home play an important part in preventing expulsion. However, pediatricians need more training in early childhood mental health and in understanding how systemic racism and implicit bias lead to preschool expulsion in children of color. By identifying children at risk for expulsion because of poverty, racial discrimination, toxic stress, insecure attachment, or history of trauma, the pediatrician can connect families with community resources that may ameliorate these effects. Pediatricians can provide information on social-emotional development in early childhood, promote positive parent-child relationships, and model and discuss age-appropriate and developmentally appropriate behavior management. Pediatricians can also guide parents toward high-quality child care programs that use mental health consultation and developmentally appropriate activities, both of which lessen the chance of child expulsion. Furthermore, behavioral health providers integrated into the medical home can provide consultation to child care providers on managing patients. These recommendations are consistent with our knowledge of early child brain development and support the current tenets of the American Academy of Pediatrics regarding the pediatrician's role in building resilience and buffering toxic stress to promote optimal child development.

Radiotherapy (RT) is an effective treatment for dogs presented with neurologic signs caused by pituitary tumors. However, its impact on the outcome of concurrent pituitary-dependent hypercortisolism (PDH) is controversial. Determine whether dogs with PDH have longer survival after pituitary RT compared with dogs with nonhormonally active pituitary masses and to evaluate whether clinical, imaging, and RT variables affect survival. Ninety-four dogs divided into 2 groups: PDH and non-PDH, based on the presence of hypercortisolism. Forty-seven dogs were allocated to the PDH group and 47 to the non-PDH group. Retrospective cohort study in which clinical records of dogs undergoing RT for pituitary macroadenomas between 2008 and 2018 at 5 referral centers were retrospectively evaluated. Survival was not statistically different between PDH and non-PDH groups (median survival time [MST], 590 days; 95% confidence interval [CI], 0-830 days and 738 days; 95% CI, 373-1103 days, respectively; P = .4). A definitive RT protocol was statistically associated with longer survival compared with a palliative protocol (MST 605 vs 262 days, P = .05). The only factor statistically associated with survival from multivariate Cox proportional hazard analysis was total radiation dose (Gy) delivered (P < .01). No statistical difference in survival was identified between the PDH and non-PDH groups, and longer survival was associated with higher Gy delivered.