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PurposeModels to study metastatic disease in rare cancers are needed to advance preclinical therapeutics and to gain insight into disease biology. Osteosarcoma is a rare cancer with a complex genomic landscape in which outcomes for patients with metastatic disease are poor. As osteosarcoma genomes are highly heterogeneous, multiple models are needed to fully elucidate key aspects of disease biology and to recapitulate clinically relevant phenotypes.Experimental designMatched patient samples, patient-derived xenografts (PDX), and PDX-derived cell lines were comprehensively evaluated using whole-genome sequencing and RNA sequencing. The in vivo metastatic phenotype of the PDX-derived cell lines was characterized in both an intravenous and an orthotopic murine model. As a proof-of-concept study, we tested the preclinical effectiveness of a cyclin-dependent kinase inhibitor on the growth of metastatic tumors in an orthotopic amputation model.ResultsPDXs and PDX-derived cell lines largely maintained the expression profiles of the patient from which they were derived despite the emergence of whole-genome duplication in a subset of cell lines. The cell lines were heterogeneous in their metastatic capacity, and heterogeneous tissue tropism was observed in both intravenous and orthotopic models. Single-agent dinaciclib was effective at dramatically reducing the metastatic burden.ConclusionsThe variation in metastasis predilection sites between osteosarcoma PDX-derived cell lines demonstrates their ability to recapitulate the spectrum of the disease observed in patients. We describe here a panel of new osteosarcoma PDX-derived cell lines that we believe will be of wide use to the osteosarcoma research community.

Ectodomain phosphatase/phosphodiesterase-1 (ENPP1) is overexpressed on cancer cells and functions as an innate immune checkpoint by hydrolyzing extracellular cyclic guanosine monophosphate adenosine monophosphate (cGAMP). Biologic inhibitors have not yet been reported and could have substantial therapeutic advantages over current small molecules because they can be recombinantly engineered into multifunctional formats and immunotherapies. Here we used phage and yeast display coupled with in cellulo evolution to generate variable heavy (VH) single-domain antibodies against ENPP1 and discovered a VH domain that allosterically inhibited the hydrolysis of cGAMP and adenosine triphosphate (ATP). We solved a 3.2 Å-resolution cryo-electron microscopy structure for the VH inhibitor complexed with ENPP1 that confirmed its new allosteric binding pose. Finally, we engineered the VH domain into multispecific formats and immunotherapies, including a bispecific fusion with an anti-PD-L1 checkpoint inhibitor that showed potent cellular activity.

BackgroundApproximately 50% of patients who receive anti-CD19 CAR-T cells relapse, and new immunotherapeutic targets are urgently needed. We recently described CD72 as a promising target in B-cell malignancies and developed nanobody-based CAR-T cells (nanoCARs) against it. This cellular therapy design is understudied compared with scFv-based CAR-T cells, but has recently become of significant interest given the first regulatory approval of a nanoCAR in multiple myeloma.MethodsWe humanized our previous nanobody framework regions, derived from llama, to generate a series of humanized anti-CD72 nanobodies. These nanobody binders were inserted into second-generation CD72 CAR-T cells and were evaluated against preclinical models of B cell acute lymphoblastic leukemia and B cell non-Hodgkin's lymphoma in vitro and in vivo. Humanized CD72 nanoCARs were compared with parental ("NbD4") CD72 nanoCARs and the clinically approved CD19-directed CAR-T construct tisangenlecleucel. RNA-sequencing, flow cytometry, and cytokine secretion profiling were used to determine differences between the different CAR constructs. We then used affinity maturation on the parental NbD4 construct to generate high affinity binders against CD72 to test if higher affinity to CD72 improved antitumor potency.ResultsToward clinical translation, here we humanize our previous nanobody framework regions, derived from llama, and surprisingly discover a clone ("H24") with enhanced potency against B-cell tumors, including patient-derived samples after CD19 CAR-T relapse. Potentially underpinning improved potency, H24 has moderately higher binding affinity to CD72 compared with a fully llama framework. However, further affinity maturation (KD

A new reading can lose effectiveness if not implemented consistently by all teachers. The purpose of this study was to understand how first grade teachers perceived implementation of the Core Knowledge Language Arts (CKLA)reading program and what challenges they encountered when using the program. Three research questions were explored in the study. RQ 1: How do first grade teachers perceive the Core Knowledge Language Arts reading program? RQ 2: What challenges do first grade teachers experience when teaching the Core Knowledge Language Arts reading program? RQ 3: How do first grade teachers implement the reading program as evidenced by their lesson plans? First grade teachers along with special education and Title 1 teachers who support first grade students were interviewed. The teachers who participated in the study had been trained in using the CKLA reading program. The interviews were transcribed into Microsoft Word documents and participants checked the transcriptions for accuracy. Through content analysis of the interviews, three themes from the interviews emerged. They were: 1) Strong Reading Skills Core, 2) Insufficient Interventions, and 3) Inadequate Writing Instruction. Dwecks mindset theory guided this study.

With the proliferation of students' abilities to use technology, in particular virtual reality (VR) applications, teachers are experimenting with VR in virtual learning environments (VLE) to engage students; but clear ethics and safety guidelines are not available. Within the field of education additional research is needed to determine how VR is employed in VLEs. Additionally, what are the decision-making processes of educators for the VR programs chosen for classroom use based on ethics and student safety? Through qualitative inquiry, educators from K-12 and higher education shared their perceptions of ethics and safety concerns in VR applications for use in the classroom. We collected data from a sample of K-12 and higher education professionals from the Association for Educational Communications and Technology (AECT) membership, an educational organization emphasizing technology integration and learning. Data analysis from the individual interviews with open-ended questions revealed five primary themes and twelve sub-themes. Participants agreed that they need ethical and safety guidelines as they embark on new horizons that influence student well-being. The findings from this study may provide an in-depth understanding of the importance of developing ethics and safety guidelines for educational VR programs when used by students.

Overlapping myelodysplastic/myeloproliferative neoplasms (MDS/MPN) are clonal hematopoietic disorders with features of myelodysplasia and myeloproliferation. The only well-characterized MDS/MPN in children is juvenile myelomonocytic leukemia, an aggressive disorder of infants and toddlers. The biochemical hallmark of this disease is hyperactivation of the Ras/MAPK signaling pathway caused by mutations in Ras pathway genes in more than 90% of patients. Translocations involving receptor tyrosine kinases have been identified in rare cases. Here, we report a 2-year-old patient who presented with MDS/MPN driven by a cytogenetically cryptic NUP98-NSD1 fusion, a translocation thought to exclusively occur in patients with acute myeloid leukemia.

The COVID-19 pandemic and the migration to online learning have presented unique challenges to undergraduate research, reducing opportunities to participate in high-impact practices, including in-person summer research programs. To mitigate some of these effects, Loyola Marymount University adapted its summer undergraduate research program, traditionally held as a six-week program on campus, to a remote structure. This article describes methods for migrating an existing program online and utilizes a mixed-methods approach to analyze student satisfaction and self-perceived changes in transferable knowledge and life skills and confidence in research ability. The data supported the hypothesis that a virtual summer research program can provide meaningful student research experiences, supportive relationships with faculty mentors, and an increase in students' desire to pursue research opportunities in the future.

BackgroundAnti-retroviral therapy (ART) regimes for HIV are associated with raised levels of circulating triglycerides (TG) in western populations. However, there are limited data on the impact of ART on cardiometabolic risk in sub-Saharan African (SSA) populations.MethodsPooled analyses of 14 studies comprising 21 023 individuals, on whom relevant cardiometabolic risk factors (including TG), HIV and ART status were assessed between 2003 and 2014, in SSA. The association between ART and raised TG (>2.3 mmol/L) was analysed using regression models.FindingsAmong 10 615 individuals, ART was associated with a two-fold higher probability of raised TG (RR 2.05, 95% CI 1.51-2.77, I2=45.2%). The associations between ART and raised blood pressure, glucose, HbA1c, and other lipids were inconsistent across studies.InterpretationEvidence from this study confirms the association of ART with raised TG in SSA populations. Given the possible causal effect of raised TG on cardiovascular disease (CVD), the evidence highlights the need for prospective studies to clarify the impact of long term ART on CVD outcomes in SSA.

Anti-retroviral therapy (ART) regimes for HIV are associated with raised levels of circulating triglycerides (TG) in western populations. However, there are limited data on the impact of ART on cardiometabolic risk in sub-Saharan African (SSA) populations. Pooled analyses of 14 studies comprising 21 023 individuals, on whom relevant cardiometabolic risk factors (including TG), HIV and ART status were assessed between 2003 and 2014, in SSA. The association between ART and raised TG (>2.3 mmol/L) was analysed using regression models. Among 10 615 individuals, ART was associated with a two-fold higher probability of raised TG (RR 2.05, 95% CI 1.51-2.77, I2=45.2%). The associations between ART and raised blood pressure, glucose, HbA1c, and other lipids were inconsistent across studies. Evidence from this study confirms the association of ART with raised TG in SSA populations. Given the possible causal effect of raised TG on cardiovascular disease (CVD), the evidence highlights the need for prospective studies to clarify the impact of long term ART on CVD outcomes in SSA.

The pediatric developmental passport was created to aid service navigation for children with autism spectrum disorder (ASD). A pilot-randomized-control trial was conducted at two developmental clinics. The intervention group received the Passport card versus the control group who received the placebo card. Primary outcome was the proportion of families who contacted ASD services 1-year following diagnosis. Of 40 families, 95% in the intervention group contacted services versus 70% in the control (p = 0.04). All families at the academic site contacted services; at the community site 90% in the intervention group contacted versus 40% in the control (p = 0.02). The Passport shows promise aiding families of children with ASD in service navigation, particularly at community clinics where specialist follow-up is not readily available.

Context: Abortion care using novel no-test telemedicine protocols and delivery of abortion pills by mail provides an historic opportunity for primary care to include medication abortion as part of the comprehensive reproductive care provided to patients. Because in-person care in the clinical setting was the only model, it is unknown whether communication practices by clinicians warrants a shift with telemedicine abortion. Objective: to evaluate best practices in patient- provider communication for telemedicine abortion care. Study Design/Analysis: We conducted in-depth interviews and used inductive-deductive constant comparative analysis. A community advisory board guided our interview guide and interpretation of findings. Setting: High-volume reproductive healthcare clinic organization with several satellite locations throughout Washington State. Population Studied: Patients who recently sought either in-clinic or telemedicine medication abortion services with licensed advance practice and family medicine providers. Instrument: Miller's conceptual framework for patient-clinician communication in telemedicine settings shaped the interview guide. Outcome Measures: Decision to choose in-clinic or telemedicine services and experiences with scheduling process, attending the appointment and communication of the clinician. Results: 30 research participants, 20 telemedicine and 10 in-clinic, completed the interview. Telehealth patients reported similar levels of interpersonal connection to their providers as in-clinic patients, including feeling their provider was relatable, supportive, affirming of their decision, not-rushed, empathetic, and an attentive listener. Some telehealth patients reported an enhanced communication experience due to their decreased anxiety and stress from avoiding an in-clinic visit. Both groups of patients understood the plan of care, including how to obtain and take medications, and felt empowered to garner personal and medical support as necessary. All patients relied heavily on printed clinic materials and online resources to answer symptomatic questions during the abortion process. Conclusions: Patients receiving either telemedicine or in-clinic abortion services by primary care providers reported similar interpersonal and medical information communication experiences. Challenging contextual factors reported by some in-clinic patients suggests telehealth as a favorable option for medication abortion care services., Presenters Emily Godfrey, MD, MPH, Anna Fiastro, MPH, MEM, Molly Ruben, MPH, Elizabeth Young, MD, MSc, Ian Bennett, MD, PhD, Elizabeth Jacob-Files, MA, [...]

Receptor tyrosine kinases (RTKs) can show extensive crosstalk, directly and indirectly. Elucidating RTK crosstalk remains an important goal in the clinical combination of anti-cancer therapies. Here, we present mass spectrometry and pharmacological approaches showing the hepatocyte growth factor receptor (MET)-promoting tyrosine phosphorylation of the epidermal growth factor receptor (EGFR) and other membrane receptors in MET-amplified H1993 NSCLC cells. Conversely, in H292 wt-EGFR NSCLC cells, EGFR promotes the tyrosine phosphorylation of MET. Reciprocal regulation of the EGFR and insulin receptor (IR) was observed in the GEO CRC cells, where inhibition of the EGFR drives tyrosine phosphorylation of the insulin receptor. Similarly, in platelet-derived growth factor receptor (PDGFR)-amplified H1703 NSCLC cells, inhibition of the EGFR promotes the tyrosine phosphorylation of the PDGFR. These RTK interactions are used to illustrate basic principles applicable to other RTK signaling networks. More specifically, we focus on two types of RTK interaction: (1) co-option of one RTK by another and (2) reciprocal activation of one receptor following the inhibition of a distinct receptor.