High-dose TSLP Induces Co-receptor Internalization and Signal Shutdown in Ph-like B-ALL with Overexpression of CRLF2

INTRODUCTION AND OBJECTIVES: Approximately half of all Ph-like B cell acute lymphoblastic leukemia is characterized by overexpression of CRLF2 (CRLF2 B-ALL). CRLF2 B-ALL is associated with high rates of relapse and is more prevalent in Hispanic children with Native American ancestry. CRLF2, together with the IL-7 receptor alpha (IL-7Ra), comprises a receptor complex that is activated by the cytokine, TSLP. Receptor activation by TSLP induces JAK2/STAT5 and PI3/AKT/mTOR signals that promote survival and proliferation of leukemia cells. To study the role of TSLP in CRLF2 B-ALL, we developed a patient-derived xenograft (PDX) model of CRLF2 B-ALL that allows us to vary circulating levels of human TSLP (hTSLP) in a physiologic environment. We generated PDX from patients’ CRLF2 B-ALL cells and compared leukemia burden in mice with varying levels of hTSLP. CRLF2 B-ALL cells grew robustly in PDX models with hTSLP levels at or below levels present in pediatric cancer patients (~10 pg/mL). In contrast, CRLF2 B-ALL cells were essentially eliminated in PDXs with hTSLP at high physiological levels (40-140 pg/mL). These data suggest that high physiologic levels of TSLP exert a potent anti-leukemia effect in Ph-like B-ALL with overexpression CRLF2. The objective of the proposed research was to evaluate potential mechanisms of TSLP’s anti-leukemia effects. METHODS: TSLP dose response studies were performed and flow cytometry was used to evaluate the effect of TSLP on CRLF2 signaling, surface expression of TSLP receptor components (IL-7Ra and CRLF2) and expression of the Suppressor of Cytokine Signaling (SOCS) proteins in CRLF2 B-ALL cell lines (MUTZ5 and CALL-4) and CRLF2 B-ALL cells from Hispanic pediatric patients. Whole genome microarray was used to evaluate TSLP effects on SOCS gene expression in patient samples. RESULTS: CRLF2 B-ALL cell lines cultured with TSLP showed a dose-dependent loss in the ability to induce phosphorylation of STAT5 and S6 (downstream of PI3/AKT/mTOR) following TSLP stimulation. This loss was correlated with a complete loss of surface IL-7Ra, and maintained for 24-48 hours following a pulse of high-dose, but not low-dose, hTSLP. The loss of surface CRLF2 was minimal. The loss of signaling and surface IL-7Ra could be prolonged if high-dose hTSLP levels were maintained. Similarly, preliminary studies of CRLF2 B-ALL cells from pediatric patients showed a loss of CRLF2 signaling and surface IL-7Ra following culture with high-dose TSLP. A potential mechanism for the effects of high-dose TSLP are the suppressor of cytokine signaling (SOCS) genes. These genes encode a family of proteins (SOCS1-7 and CISH) that regulate cytokine signaling via negative feedback through multiple mechanisms including ubiquitin-mediated cytokine receptor degradation. Whole genome microarray showed that SOCS1, SOCS2, SOCS3 and CISH mRNA are upregulated in patient CRLF2 B-ALL cells cultured with high-dose TSLP. Flow cytometry analysis showed that high-dose TSLP upregulates SOCS1 and SOCS3 proteins in CRLF2 B-ALL cell lines and in patient samples. CONCLUSIONS: These data provide evidence that TSLP exerts anti-leukemia effects by shutting down CRLF2-mediated signals and that these effects are at least partially mediated by the loss of the IL-7Ra component, and potentially through SOCS family proteins. These studies identify the human TSLP cytokine as a potential biologic therapy to treat CRLF2 B-ALL

Musical Borrowing in Las Cuatro Estaciones Portenas: Piazzolla, Desyatnikov, Vivaldi

Leonid Desyatnikov arranged Astor Piazzolla\u27s Las Cuatro Estaciones Portenas for violin and string orchestra, interspersing quotations from Vivaldi\u27s Four Seasons throughout the new work. My score-based analysis of the arrangement investigates Desyatnikov\u27s borrowing practice and connects his compositional techniques to musicological meanings. In addition to identifying levels of contrast value between the quotation and the surrounding arrangement, I examine alterations to the original content and context of the quotations and determine whether Desyatnikov maintains or transforms the quotations\u27 identities. I combine identity and contrast value into an interpretive model of musical interaction, providing a more nuanced exploration of musical dominance

P-60 Let All the People Praise Thee: Early Lutheran Chorales and the Priesthood of All Believers

Martin Luther articulated his belief in the priesthood of all believers through both word and music. His chorales reflect his desire for direct congregational participation in the worship service, leading to personal experiences of justification by faith when people met their true High Priest without an earthly mediator. Luther used the chorale as a tool for teaching theology and an occasion of an individual encounter with God. I demonstrate the chorale’s role as a tool for understanding and experiencing justification by faith, by studying the theme of joy in selected examples from the hymnody of Luther and his contemporaries

Every-Other-Day Clutch-Initiation Synchrony as an Adaptive Response to Egg Cannibalism in Glaucous-Winged Gulls (Larus glaucescens)

Sea surface temperatures (SST) in the Pacific Northwest of North America rose 1 °C during the last half of the twentieth century. Cannibalism, a behavior observed in diverse taxa, is often associated with low food supplies, which for marine animals can be precipitated by high SSTs. In an 8 year study, we found that in years of higher sea surface temperatures, Glaucous-winged Gulls (Larus glaucescens) breeding in a colony in Washington State, Salish Sea, USA, tended to exhibit higher rates of egg cannibalism by males, higher levels of every-other-day clutch-initiation synchrony by females, and longer egg-laying seasons than in years of lower SST. Clutch-initiation synchrony increased the odds that an egg survived cannibalism and may serve as an adaptive response to egg cannibalism. Short-term climate and resource fluctuations associated with El Niño Southern Oscillation (ENSO) events may select for behavioral plasticity in marine organisms, allowing long-lived individuals such as marine birds to switch between alternative life history tactics. The implications for long-term SST warming, however, remain unknown

Data for Henson et al. 2016

These data are archived for the paper Adaptation to short-term ENSO fluctuations may provide tipping points for populations subjected to long-term climate change by Henson et al., submitted. Additional files: Supplementary Figure 1 | Sample plots on Violet Point, Protection Island National Wildlife Refuge, Washington, USA. The larger colony extends throughout much of the spit