Gene fusions prevalent in prostate cancer (CaP) lead to the elevated expression of the ERG proto-oncogene. ERG activation present in 50–70% of prostate tumors underscores one of the most common oncogenic alterations in CaP. Despite numerous reports of gene fusions and mRNA expression, ERG oncoprotein status in CaP still remains to be defined. Furthermore, development of ERG protein-based assays may provide a new dimension to evaluation of gene fusions involving diverse androgen-regulated promoters and the ERG protein-coding sequence. Through exhaustive evaluations of 132 whole-mount prostates (261 tumor foci and over 200 000 benign glands) for the ERG oncoprotein nuclear expression, we demonstrated 99.9% specificity for detecting prostate tumor cells using a highly specific anti-ERG monoclonal antibody. The ERG oncoprotein expression correlated well with fusion transcript or gene fusion in randomly selected specimens. Strong concordance of ERG-positive foci of prostatic intraepithelial neoplasia (PIN) with ERG-positive carcinoma (82 out of 85 sections with PIN, 96.5%) affirms the biological role of ERG in clonal selection of prostate tumors in 65% (86 out of 132) of patients. Conversely, ERG negative PINs were associated with ERG-negative carcinoma. Taken together, the homogeneous and strong ERG expression detected in individual tumors establishes the potential for ERG oncoprotein-based stratification of CaP.

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ERG oncoprotein expression in prostate cancer: clonal progression of ERG-positive tumor cells and potential for ERG-based stratification

Purpose: Gene expression studies in melanoma have been few because tumors are small and cryopreservation is rarely possible. The purpose of this study was to evaluate the Illumina DASL Array Human Cancer Panel for gene expression studies in formalin-fixed melanoma primary tumors and to identify prognostic biomarkers. Experimental Design: Primary tumors from two studies were sampled using a tissue microarray needle. Study 1: 254 tumors from a melanoma cohort recruited from 2000 to 2006. Study 2: 218 tumors from a case-control study of patients undergoing sentinel node biopsy. Results: RNA was obtained from 76% of blocks; 1.4% of samples failed analysis (transcripts from SPP1 ; P = 2.11 × 10 −6 ) and supportive evidence for this was obtained in study 2 used as a validation set ( P = 0.006; unadjusted data). Osteopontin level in study 1 remained a significant predictor of relapse-free survival when data were adjusted for age, sex, tumor site, and histologic predictors of relapse. Genes whose expression correlated most strongly with osteopontin were PBX1, BIRC5 (survivin), and HLF . Conclusion: Expression data were obtained from 74% of primary melanomas and provided confirmatory evidence that osteopontin expression is a prognostic biomarker. These results suggest that predictive biomarker studies may be possible using stored blocks from mature clinical trials. (Clin Cancer Res 2009;15(22):6939–46)

https://clincancerres.aacrjournals.org/content/clincanres/15/22/6939.full.pdf

Gene expression profiling of paraffin-embedded primary melanoma using the DASL assay identifies increased osteopontin expression as predictive of reduced relapse-free survival.

Purpose:
Enhanced tumor cell survival through expression of inhibitors of apoptosis (IAP) is a hallmark of cancer. Survivin, an IAP absent from most normal tissues, is over-expressed in many malignancies and associated with a poorer prognosis. We report the First-in-Human dose study of LY2181308, a second generation antisense oligonucleotide (ASO) directed against survivin mRNA. 
Experimental Design:
A dose-escalation study evaluating the safety, pharmacokinetics and pharmacodynamics of LY2181308 administered intravenously for 3-hours as a loading dose on three consecutive days and followed by weekly maintenance doses. Patients were eligible after signing informed consent, had exhausted approved anti-cancer therapies and agreed to undergo pre- and post-treatment tumor biopsies to evaluate reduction of survivin protein and gene expression. 
Results:
A total of 40 patients were treated with LY2181308 at doses of 100 to 1000 mg. Twenty-six patients were evaluated at the recommended Phase 2 dose of 750 mg, at which level serial tumor sampling and [11C]LY2183108 PET imaging demonstrated that ASO accumulated within tumor tissue, reduced survivin gene and protein expression by 20% and restored apoptotic signaling in tumor cells in vivo. Pharmacokinetics were consistent with pre-clinical modeling, exhibiting rapid tissue distribution and terminal half-life of 31 days. 
Conclusions:
The tumor-specific, molecularly targeted effects demonstrated by this ASO in man underpin confirmatory studies evaluating its therapeutic efficacy in cancer.

https://clincancerres.aacrjournals.org/content/clincanres/16/24/6150.full.pdf

Tumor Survivin is Downregulated by the Antisense Oligonucleotide LY2181308: A Proof of Concept, First-in-Human Dose Study

Materials and methods related to diagnosing and/or determining prognosis of prostate cancer.

Materials and methods for determining diagnosis and prognosis of prostate cancer

Aims

Synaptic vesicle proteins 2 (SV2) are neuronal vesicles membrane glycoproteins that appear as important targets in the treatment of partial and generalized epilepsies. Therefore, we analysed the expression of SV2 isoforms in the hippocampus of patients with temporal lobe epilepsy (TLE).



Methods

SV2A, SV2B and SV2C immunostaining and QuantiGene branched DNA assay were performed on biopsies from 31 consecutive TLE patients with mesial temporal sclerosis (MTS) and compared with 10 autopsy controls. SV2 expression was further compared with Timm's staining, and synaptophysin, Zinc transporter 3 (ZnT3), dynorphin, vesicular glutamate transporter 1 (VGLUT1) and vesicular GABA transporter (VGAT) expression.



Results

In TLE patients, SV2A and SV2B expression was decreased in areas of synaptic loss. SV2C, which is weakly expressed or absent in the hippocampus of controls, was overexpressed in 10/11 cases with classical MTS1A and mossy fibre sprouting but not in cases with other types of MTS. SV2C staining was located in the inner molecular layer of the dentate gyrus and colocalized with dynorphin, ZnT3 and VGLUT1, suggesting selective expression in presynaptic glutamatergic Zn2+-rich terminals of abnormal sprouting fibres. SV2 expression patterns correlated with histological subtypes of MTS, but not with clinical features or therapeutic regimens in this patient cohort.



Conclusion

In classical MTS1A, the expression of SV2 isoforms is altered with a marked decrease of SV2A and SV2B paralleling synaptic loss and a selective increase of SV2C in sprouting mossy fibres. These findings suggest a different physiology of sprouting synapses and the possibility to target them with SV2C-specific strategies.

Expression pattern of synaptic vesicle protein 2 (SV2) isoforms in patients with temporal lobe epilepsy and hippocampal sclerosis

Evaluation of the branched-chain DNA assay for measurement of RNA in formalin-fixed tissues

Background Faculty development (FD) encompasses structured programming that aims to enhance educator knowledge, skill, and behavior. No uniform framework for faculty development exists, and academic institutions vary in their faculty development programming, ability to overcome barriers, resource utilization, and achievement of consistent outcomes. Objective The authors aimed to assess current FD needs among emergency medicine educators from six geographically and clinically distinct academic institutions to inform overall faculty development advancement in emergency medicine (EM). Methods This cross-sectional study assessed FD needs among EM educators. A survey was developed, piloted, and distributed to faculty via each academic institution's internal e-mail listserv. Respondents were asked to rate their comfort level with and interest in several domains of FD. Respondents were also asked to identify their previous experience, satisfaction with the FD they have received, and barriers to receiving FD. Results Across six sites, 136/471 faculty completed the survey in late 2020 (response rate of 29%): 69.1% of respondents reported being satisfied overall with the FD they have participated in, and 50.7% reported being satisfied with education FD specifically. Faculty report higher comfort levels and interest in several domains when satisfied with the education-specific FD they have received compared with those who report not being satisfied. Conclusions EM faculty report generally high satisfaction with the overall faculty development they have received, although only half express satisfaction with their education-related faculty development. Faculty developers in EM may incorporate these results to inform future faculty development programs and frameworks. 

Jonathan Karademos

Papers

Evaluation of the branched-chain DNA assay for measurement of RNA in formalin-fixed tissues

Faculty Development and the Emergency Medicine Educator: A National Needs Assessment.