Evaluation of optical coherence tomography findings in age-related macular degeneration: a reproducibility study of two independent reading centres

International audienceBackground/aims : To determine the reproducibility among readers of two independent certified centers, the Vienna Reading Center (VRC) and the University of Wisconsin-Madison Reading Center (UW-FPRC) for OCT images in age-related macular degeneration (AMD). Methods : Fast macular thickness scans and 6 mm cross hair scans were obtained from 100 eyes with all subtypes of AMD using Stratus OCT. Consensus readings were performed by two certified OCT readers of each Reading Center using their grading protocol. Common variables of both grading protocols, such as presence of cystoid spaces, subretinal fluid, vitreomacular traction and retinal pigment epithelial detachment were compared using kappa statistics. In addition, the intraclass correlation coefficient (ICC) was calculated for center point thickness (CPT) of values remeasured manually in the presence of alignment errors. Results : The reproducibility was dependent on the variable measured with a kappa value of 0.81 for the presence of cystoid spaces, 0.78 for the presence of subretinal fluid and 0.795 for the presence of vitreomacular traction. The lowest reproducibility was found for the presence of retinal pigment epithelial detachment with a kappa value of 0.51. The CPT was remeasured in 29 out of 100 scans at both sites with an ICC of the remeasured thicknesses of 0.92. Conclusion : OCT scan data are crucial in monitoring treatment efficacy in AMD clinical trials. For comparison of results obtained by different Reading Centers, the inter-Reading Center reproducibility is essential. Although the reproducibility is generally high, the reliability depends on the selected morphological parameters

Beyond the Individual: Sexual Minority Help-Seeking and the Consequences of Structural Barriers

Sexual minorities report more psychological distress, unmet mental health needs, and barriers to mental health care compared to heterosexuals, yet little is known about their barriers to seeking out mental health care. The present study reports the factors that influence intentions to seek out mental health care of a national survey of 398 sexual minorities. Structural equation modeling identified structural barriers, such as cost, time, and knowing how to access services, as the strongest predictor of sexual minorities’ help-seeking intentions. Latent moderators indicated sexual minorities’ help-seeking intentions varied depending upon their degree of psychological distress. This revealed a pattern where the most vulnerable participants (i.e., those with high structural barriers and negative help-seeking attitudes) were willing to pursue mental health care only when they were experiencing significant distress. Further, nearly 40% of participants reported unmet mental health needs, and structural barriers were the primary reasons for this deficit. Findings from this study contrast with previous mental health help-seeking research by emphasizing the importance of structural vulnerability, which refers to the external forces that frame and constrain choices, thereby impeding decision-making and limiting life options for those who are in systemically disadvantaged social positions. These findings are discussed in terms of counseling psychology training, practice, social justice advocacy and future healthcare research

Relationship quality and the mentoring of aggressive, high-risk children

We used data from a randomized clinical trial to examine the degree to which relationship quality predicted outcomes for aggressive children in two different mentoring programs. Data were available for 145 aggressive children in Grades 2 and 3. Children were blocked by school and randomly assigned to PrimeTime (n ¼ 75) or Lunch Buddy (n ¼ 70) programs. PrimeTime combined community-based mentoring with child-focused skills training and consultation for parents and teachers, and mentors were extensively trained and supervised. Lunch Buddy was a stand-alone, school-based mentoring program that involved lunchtime visits and a different mentor each semester. PrimeTime children rated their mentors as more supportive than did Lunch Buddy children. Relationship conflict predicted changes in teacher-rated externalizing problems. Ratings of relationship quality interacted with treatment in predicting changes in parent-rated externalizing behavior for PrimeTime children only

Abstract 2654: Inhibition of lymphoma interactions withangiogenic and lymphangiogenic endotheliumusing αCD20-IgG1-huEndoP125A, an anti-CD20 endostatin fusion protein

Abstract Mantle Cell Lymphoma (MCL) is an aggressive non-Hodgkin’s lymphoma that frequently metastasizes. Lymphoma dissemination into distant sites is the primary cause of cancer mortality. Lymphoma metastasis requires development of supporting vasculature in which the interaction between malignant B cells and the vasculature or lymphatics is critical to tumor cell growth and dissemination. Human umbilical vein endothelial cells (HUVEC) or human lymphatic endothelial cells (HLEC) cultured on Matrigel, in vitro, assemble into networks of capillary like structures (CLS) or lymphatic like structures (LLS). When Mino or Jeko-1 MCL cells are co-cultured with vascular or lymphatic cells they migrate toward and anneal to nascent CLS or LLS. We constructed a novel antibody fusion protein, αCD20-IgG1-huEndoP125A, to inhibit such interactions. αCD20-IgG1-huEndoP125A links an αCD20 human IgG1 targeting domain and a mutant version of human endostatin, huEndoP125A, with enhanced anti-angiogenic properties. αCD20-IgG1-huEndoP125A completely inhibits CLS or LLS formation and prevents the alignment and annealing of MCL to CLS or LLS. αCD20-IgG1-huEndoP125A also significantly reduces both endothelial and MCL migration and invasion in vitro. Cell migration and trafficking is tightly regulated by chemokine interactions with corresponding chemokine receptors. We assayed for the presence of chemokines using a dot blot chemokine array. Jeko-1 and HUVEC cells were cultured on Matrigel, alone or together in co-culture, and either treated with αCD20-IgG1-huEndoP125A or left untreated. CXCL12 was expressed at high levels in HUVEC cultures undergoing CLS formation and expression was further increased in HUVEC and MCL co-cultures. HUVEC or HUVEC/MCL co-cultures treated with αCD20-IgG1-huEndoP125A showed markedly decreased CXCL12 expression. Flow cytometry showed that corresponding chemokine receptor CXCR4 was upregulated on MCL cells compared to normal B cells. CXCR4 was also upregulated on HUVEC undergoing CLS formation, and further increased on HUVEC in co-culture with MCL cells. αCD20-IgG1-huEndoP125A inhibited HUVEC CLS formation, reduced MCL migration to and alignment with nascent CLS or LLS, downregulated CXCL12 levels in HUVEC or coculture supernatants, and reduced CXCR4 expression on both HUVEC and MCL cells. Murine 38c13-huCD20 lymphoma cells were implanted s.c. into C3H mice and mice were treated i.p. with either PBS, Rituximab, or equimolar αCD20-IgG1-huEndoP125A. Mice treated with αCD20-IgG1-huEndoP125A showed significantly reduced tumor growth compared to mice treated with PBS or rituximab. αCD20-IgG1-huEndoP125A ability to inhibit angiogenesis, lymphangiogenesis, and lymphoma association with angiogenic or lymphatic vasculature may be a potent means of preventing lymphoma growth and metastasis. Citation Format: Christian Elledge, Seung-Uon Shin, Rathin Das, Yu Zhang, Hyun-Mi Cho, Sundaram Ramakrishnan, Ankita Sankar, Joseph Rosenblatt. Inhibition of lymphoma interactions withangiogenic and lymphangiogenic endotheliumusing αCD20-IgG1-huEndoP125A, an anti-CD20 endostatin fusion protein [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2654

Abstract 6319: Mechanism of anti-EGFR antibody endostatin fusion protein action on inhibition of vasculogenic mimicry and tumor cell motility in triple negative breast cancer

Abstract Triple Negative Breast Cancer (TNBC) is an aggressive subtype of breast cancer with high metastatic potential and increased morbidity and mortality. Although expression of and signaling by the epidermal growth factor receptor (EGFR) is commonly seen in TNBC, anti-EGFR antibodies such as Cetuximab have had limited therapeutic efficacy, used alone or in combination with chemotherapy. Primary TNBC tumor growth and metastases require supporting vasculature, which develops through a combination of endothelial angiogenesis and vasculogenic mimicry (VM). VM is frequently seen in TNBC and is associated with aggressive metastatic behavior. We previously developed αEGFR-E-P125A, an antibody-endostatin fusion protein, linking an anti-EGFR antibody to a mutated version of the anti-angiogenic protein endostatin (E-P125A). αEGFR-E-P125A delivers a dimeric E-P125A payload which inhibits TNBC angiogenesis and VM in vitro and in vivo, and markedly decreases metastasis in multiple TNBC xenograft models. To study changes in the transcriptome following induction of VM and inhibition of VM with αEGFR-E-P125A, RNA-seq was conducted on MDA-MB-231-4175 TNBC cells grown in a 2D monolayer and compared to TNBC cells undergoing VM when plated in matrigel. A second comparison was made between TNBC cells undergoing VM and TNBC cells in which VM was inhibited by treatment with αEGFR-E-P125A. Differential gene expression analysis and gene set enrichment analysis (GSEA), were used to define pathways which were inversely regulated during the induction of VM on matrigel and inhibition of VM with αEGFR-E-P125A. Gene set enrichment analysis demonstrated that αEGFR-E-P125A treatment significantly downregulated genes in the JAK-STAT and angiogenesis signaling pathways. Phospho-array analysis demonstrated decreased EGFR phosphorylation at Y1069 in addition to decreased phosphorylation of FAK Y397 and STAT3 Y705 sites downstream of α5β1 integrin. Since inhibition of EGFR signaling alone did not inhibit VM, and endostatin is a known ligand of α5β1 integrin, we focused on effects of αEGFR-E-P125A on inhibition of α5β1 integrin/FAK signaling pathway. Independent siRNA knockdowns of α5 integrin and FAK in MDA-MB-231-4175 cells confirmed that downregulation of α5 integrin/FAK signaling inhibited VM in vitro. αEGFR-E-P125A additionally prevented EGF-induced EGFR phosphorylation and fibronectin-induced FAK phosphorylation by α5β1 integrin. Treatment of TNBC cells with αEGFR-E-P125A reduced total α5 integrin protein levels and decreased co-localization of EGFR and α5β1 integrin receptors. These results indicate that αEGFR-E-P125A suppressed both EGFR and α5β1 integrin signaling. Simultaneous inhibition of EGFR and α5β1integrin signaling by αEGFR-E-P125A fusion is a promising approach to inhibition of TNBC growth and metastases. Citation Format: Ankita Sankar, Hava Gil Henn, Hyun Mi Cho, Dania Nassar, Sundaram Ramakrishnan, Yu Zhang, Christian Elledge, Seung Uon Shin, Joseph Rosenblatt. Mechanism of anti-EGFR antibody endostatin fusion protein action on inhibition of vasculogenic mimicry and tumor cell motility in triple negative breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 6319

Screening of microRNAs controlling body fat in Drosophila melanogaster and identification of miR-969 and its target, Gr47b.

MicroRNAs (miRNAs) are small non-protein coding RNAs and post-transcriptionally regulate cellular gene expression. In animal development, miRNAs play essential roles such as stem cell maintenance, organogenesis, and apoptosis. Using gain-of-function (GOF) screening with 160 miRNA lines in Drosophila melanogaster, we identified a set of miRNAs which regulates body fat contents and named them microCATs (microRNAs Controlling Adipose Tissue). Further examination of egg-to-adult developmental kinetics of selected miRNA lines showed a negative correlation between fat content and developmental time. Comparison of microCATs with loss-of-function miRNA screening data uncovered miR-969 as an essential regulator of adiposity. Subsequently, we demonstrated adipose tissue-specific knock-down of gustatory receptor 47b (Gr47b), a miR-969 target, greatly reduced the amount of body fat, recapitulating the miR-969 GOF phenotype