Interest in biology education research (BER) has been growing over the last two decades, yet few BER publications focus on community colleges, which serve a large percentage of the undergraduate student population and a majority of those students who identify with historically underserved groups. In this paper, we define community college biology education research (CC BER) as publications with a community college faculty member as an author, publications with a community college study context or a focus on community college biology teaching and learning, and publications that use community college students as a source of data. We conducted a literature review to quantify how CC BER has progressed since initial calls for broadening participation by recording the number of CC BER publications in seven prominent journals between 2016 and 2020. Our formal analysis of peer-reviewed BER literature indicates that there has been a statistically significant increase in CC BER publications from 3.2% to 5.9% of total BER publications since the last analysis in 2017. We conclude with a discussion of strategies for further broadening of participation in CC BER.

In March 2020, the COVID-19 pandemic altered instructional and learning strategies at institutions across the globe. This emergency transition to remote instruction (ETRI) resulted in ambiguity regarding what to teach, how to teach, and instructor/student workload.

MET is frequently overexpressed in head and neck squamous cell carcinoma (HNSCC) and degraded by c-CBL E3-ubiquitin ligase. We investigated genetic variations of c-CBL in HNSCC and the relationship between c-CBL and MET expression. High MET, low c-CBL expression was detected in 10 cell lines and 73 tumor tissues. Two novel mutations (L254S, L281F), and the single nucleotide polymorphism (SNP) P782L were identified from archival tumor tissues. 27.3% of loss of heterozygosity was found at CBL locus. Ectopic expression of wild-type c-CBL in SCC-35 cells downregulated MET expression and decreased cell viability. These results suggest MET overexpression is related to altered c-CBL expression, which may influence tumorigenesis.

Abstract Purpose/Objective(s): Casitas B-lineage lymphoma (CBL) is an E3 ubiquitin ligase and adaptor molecule that is important in cancer. It belongs in the family with CBL-B and CBL-3. Our previous studies detected CBL mutations, loss of heterozygosity, and low protein expression in non-small cell lung cancer (NSCLC). We also determined the genetic variations of CBL, their relationship to receptor tyrosine kinases (RTK) such as EGFR and MET, and their functionality in NSCLC. However, ubiquitination of EGFR showed no difference between CBL wild type (WT) and mutants (Mts). Therefore, we undertook to investigate whether MET is a potential target of CBL and better therapeutic strategy. Materials/ Methods: CBL alterations were determined in NSCLC patients with next generation sequencing. Immunoprecipitation was performed to detect the ubiquitination of MET by CBL WT and Mts. MET inhibitor SU11274 was utilized to compare cell viability of CBL WT cells with viability of CBL Mt cells. Cell motility after SU11274 treatment was examined by wound healing assay. Soft agar assay was performed to investigate colony formation of shRNA knockdown (sh-CBL) cells and SU11274 treatment. PamGene Chip analysis was used to detect and predict the RTK phosphorylation difference between CBL WT and Mt/sh-CBL cells. In vivo mouse study was performed to examine tumor growth and metastasis of sh-CBL cells. Results: CBL had 6% (8/135) mutation rate in NSCLC patient population. When CBL and MET protein expression levels were analyzed by immunoblot, CBL WT cells had lower MET protein expression than CBL Mt cells. The ubiquitination of MET was decreased in cells that transiently expressed CBL Mt relative to CBL WT cells. In addition, CBL Mt cells were more sensitive to MET inhibitor SU11274 than CBL WT cells. Wound healing assay demonstrated sh-CBL cells had an increase in cell motility and more sensitive with SU11274 migration inhibition. Furthermore, soft agar assay showed an increase in colony number and size of sh-CBL cells compared to control cells. After treatment with SU11274, sh-CBL cells showed inhibition in colony formation. In vivo study showed sh-CBL cells had lower tumor growth but more metastasis than control cells. PamGene results predicted an effective cell signaling pathway involving MET, paxillin, EPHA2, and VEGFR.. Conclusions: Ubiquitination assay result suggested MET is one of direct targets of CBL. The cell viability and motility results demonstrated that CBL mutants and sh-CBL cells are more sensitive than CBL WT cells to MET inhibitor SU11274, suggesting they have higher MET protein expression. Moreover, the PamGene results indicated that MET is involved in CBL cell signaling pathway. Thus, CBL gene status could be a potential indication for lung cancer therapy utilizing MET and other inhibitors. Citation Format: Yi-Hung Carol Tan, Cleo Rolle, Li Zhu, Minu K. Srivastava, Sherven Sharma, Ravi Salgia. Efficacy of MET inhibitors in NSCLC with CBL alterations. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4361. doi:10.1158/1538-7445.AM2015-4361

Small cell lung cancer (SCLC) is a devastating disease, and current therapies have not greatly improved the 5-year survival rates. Topoisomerase (Top) inhibition is a treatment modality for SCLC; however, the response is short lived. Consequently, our research has focused on improving SCLC therapeutics through the identification of novel targets. Previously, we identified MNNG HOS transforming gene (MET) to be overexpressed and functional in SCLC. Herein, we investigated the therapeutic potential of combinatorial targeting of MET using SU11274 and Top1 using 7-ethyl-10-hydroxycamptothecin (SN-38). MET and TOP1 gene copy numbers and protein expression were determined in 29 patients with limited (n = 11) and extensive (n = 18) disease. MET gene copy number was significantly increased (>6 copies) in extensive disease compared with limited disease (P = 0.015). Similar TOP1 gene copy numbers were detected in limited and extensive disease. Immunohistochemical staining revealed a significantly higher Top1 nuclear expression in extensive (0.93) versus limited (0.15) disease (P = 0.04). Interestingly, a significant positive correlation was detected between MET gene copy number and Top1 nuclear expression (r = 0.5). In vitro stimulation of H82 cells revealed hepatocyte growth factor (HGF)–induced nuclear colocalization of p-MET and Top1. Furthermore, activation of the HGF/MET axis enhanced Top1 activity, which was abrogated by SU11274. Combination of SN-38 with SU11274 dramatically decreased SCLC growth as compared with either drug alone. Collectively, these findings suggest that the combinatorial inhibition of MET and Top1 is a potentially efficacious treatment strategy for SCLC. Mol Cancer Ther; 13(3); 576–84. ©2013 AACR.

INTRODUCTION:Altered expression of MUC4 plays an oncogenic role in various cancers, including pancreatic, ovarian, and breast. This study evaluates the expression and role of MUC4 in non-small-cell lung cancer (NSCLC).METHODS:We used a paired system of MUC4-expressing (H292) and MUC4-nonexpressing (A549) NSCLC cell lines to analyze MUC4-dependent changes in growth rate, migration, and invasion using these sublines. We also evaluated the alterations of several tumor suppressor, proliferation, and metastasis markers with altered MUC4 expression. Furthermore, the association of MUC4 expression (by immunohistochemistry) in lung cancer samples with patient survival was evaluated.RESULTS:MUC4-expressing lung cancer cells demonstrated a less proliferative and metastatic phenotype. Up-regulation of p53 in MUC4-expressing lung cancer cells led to the accumulation of cells at the G2/M phase of cell cycle progression. MUC4 expression attenuated Akt activation and decreased the expression of Cyclins D1 and E, but increased the expression of p21 and p27. MUC4 expression abrogated cancer cell migration and invasion by altering N- & E-cadherin expression and FAK phosphorylation. A decrease in MUC4 expression was observed with increasing tumor stage (mean composite score: stage I, 2.4; stage II, 1.8; stage III, 1.4; and metastatic, 1.2; p = 0.0093). Maximal MUC4 expression was associated with a better overall survival (p = 0.042).CONCLUSION:MUC4 plays a tumor-suppressor role in NSCLC by altering p53 expression in NSCLC. Decrease in MUC4 expression in advanced tumor stages also seems to confirm the novel protective function of MUC4 in NSCLC.

Many skin infections are caused by Staphylococcus aureus, a bacterial pathogen that produces virulence factors associated with these conditions such as exfoliative toxins A and B (ETA, ETB) and the leukotoxin Panton-Valentine leukocidin (PVL). Herein, we examine the potential of skin-infecting S. aureus to produce virulence factors and their impact on the local immune response. Toxin gene profiles were generated from 188 S. aureus isolated as single infecting organisms from skin lesions and demonstrated a higher potential to express ETA, ETB, and PVL than community isolates (p < 0.001). Within the study isolate group, the prevalence of genes encoding PVL was higher among methicillin-resistant S. aureus (MRSA; n = 49), while genes encoding ETs were more prevalent in methicillin-susceptible S. aureus (MSSA; n = 139). When lesion-associated white blood cell (WBC) counts were dichotomized into high- or low-WBC-count-associated bacteria, the gene for ETA was found to be associated with a low WBC count among MSSA (p = 0.001). The ETA-induced mouse model of staphylococcal scalded skin syndrome was used to investigate the link between ETA and cytokine production. Elevated IL-6 levels in the serum and increased expression of IL-6 mRNA in the skin were detected in response to ETA exposure. These findings were recapitulated in vitro using primary human keratinocytes. Thus, S. aureus may influence the local immune response via ETA cleavage of desmoglein 1 and the induction of cutaneous IL-6 expression.

Despite progress in locoregional and systemic therapies, patient survival from lung cancer remains a challenge. Receptor tyrosine kinases are frequently implicated in lung cancer pathogenesis, and some tyrosine kinase inhibition strategies have been effective clinically. The EphB4 receptor tyrosine kinase has recently emerged as a potential target in several other cancers. We sought to systematically study the role of EphB4 in lung cancer. Here, we demonstrate that EphB4 is overexpressed 3-fold in lung tumors compared to paired normal tissues and frequently exhibits gene copy number increases in lung cancer. We also show that overexpression of EphB4 promotes cellular proliferation, colony formation, and motility, while EphB4 inhibition reduces cellular viability in vitro, halts the growth of established tumors in mouse xenograft models when used as a single-target strategy, and causes near-complete regression of established tumors when used in combination with paclitaxel. Taken together, these data suggest an important role for EphB4 as a potential novel therapeutic target in lung cancer. Clinical trials investigating the efficacy of anti-EphB4 therapies as well as combination therapy involving EphB4 inhibition may be warranted.

e17549 Background: MUC4 has been shown to play a key role in breast and pancreatic cancer. MUC4 expression is seen in the normal bronchial epithelium, but its role in non-small cell lung cancer (NSCLC) is unclear. Here we study the association between MUC4 overexpression and clinical features of NSCLC. Methods: The diagnosis of lung cancer was confirmed in H&E sections. MUC4 level was semi-quantified by immunohistochemistry. Each sample was given a composite score based on intensity and extent of staining. The intensity was graded on a 4 point scale: – (0), + (1), ++ (2), and +++ (3). The extent of staining too was graded on a 4 point scale: 1 (1-25%), 2 (26-50%), 3 (51-75%), and 4 (76-100%). The composite score was obtained by multiplying the two values, with possible scores of 0-12. Poisson mixed effects models were used to compare the score between groups. Kaplan-Meier method was used to estimate survival distributions and the log-rank test was used to compare them. Wilcoxon sign rank test was used to compare the difference in score between lung tissue and corresponding lymph node. P value <0.05 was considered to be statistically significant. Results: 167 samples from 54 non-metastatic NSCLC patients were analyzed. Of these, 13 (25%) patients had stage I, 33 (63%) had stage II and 6 (12%) patients had stage III disease. Distribution of the composite score differed significantly by stage (p=0.0043). A higher proportion of patients with stage I NSCLC had a score of 12 (77%) compared to stage II (18%) and stage III (57%) (p=0.0001). When patients who had data for both the primary tumor and corresponding metastatic lymph node were analyzed, MUC4 expression was higher in the primary tumor compared to the metastasis (p=0.05). The effect of MUC4 expression on survival was analyzed in 29 patients with survival data. Of these, 16 had a composite score of 12, while 13 had a score <12. Patients with a score <12 had significantly worse survival than those with a score of 12 (p<0.045). Conclusions: MUC4 expression is more common in early stage NSCLC and appears to be associated with a better outcome in patients with non-metastatic NSCLC. The therapeutic implications of this finding need to be investigated further.

Abstract MET is a receptor tyrosine kinase that plays a critical role in proliferation, angiogenesis, invasion, and metastasis. It has been shown to be dysregulated in a number of malignancies, including non-small cell lung cancer (NSCLC). Despite reports of alterations in MET, and its ligand, HGF, being highly associated with advanced pathological stage and worse outcome in patients with NSCLC, the value of MET as a prognostic biomarker remains unclear. The primary objective of this study is to determine the correlation between MET alterations and expression with stage and overall survival in adenocarcinoma (AC) patients in a large cohort of patients. The secondary objectives are to determine the correlation between overall survival and the following: 1) epithelial mesenchymal transition (EMT), 2) EGFR mutations & expression, 3) KRAS mutations, 4) TP53 mutations, and 5) CBL mutations. In addition, sera levels of circulating MET and HGF will be evaluated in order to determine their potential as prognostic factors. MET, EGFR exons 18–21, TP53 exons 4–10, KRAS exon 2, and CBL exons 2–16 were sequenced using standard PCR and sequencing techniques. Standard immunohistochemistry (IHC) techniques were used to evaluate MET, phosphorylated (pMET Y1003 and Y1230/34/35) p53, HGF, EGFR, and E-cadherin expression. A total of 280 patients will be included in this study, and the interim analysis reported herein evaluated 100 patients. The intensity of cytoplasmic or membranous staining was scored on fourpoint scale: (0, no staining; 1+, weak; 2+, moderate; 3+, strong staining). The extent of staining was scored on a similar scale (0, negative; 1+, 1–10%; 2+, 11–50%; 3+, > 50%). The product of the intensity and extent of staining yielded final scores between 0 and 9. The mean expressions were: MET 3.6 (±0.3); pY1230/34/35 MET 2.0 (±0.2); pY1003MET 4.6 (±0.3); HGF 4.4 (±0.3); EGFR 4.3 (±0.3); TP53 3.7 (±0.3); and E-cadherin 5.5 (±0.3). In ten patient samples, six non-synonymous (NS) mutations were detected in MET (SEMA domain: E168D, M362T, N375S, and Q318K; JM domain: T992I and R970C). In EGFR, the NS mutation L858R was detected in two patients. We detected 12 NS mutations in TP53 (exon 4: E68*, P72R; exon 5: V157F, R175H, I162F, H193Y, Y163D; exon 8: R273L, R273C, V274L, A276F, and G266*). Four NS mutations were detected in exon 2 of KRAS (G12C, G12V, G12D, and G12S). Three mutations were found in the proline-rich region of CBL (L676P, A677S and A678S). ELISAs were utilized to determine soluble MET and HGF levels in pre- and postoperative sera samples. Soluble MET serum levels were significantly increased (p<0.0005) in post-operative samples (1760 ng/ml ± 51.66) compared to pre-operative samples (1585 ng/ml ± 44.61). HGF levels were similar in pre-operative (1008 pg/ml ± 74.8) and post-operative samples (1266 pg/ml ± 175.9). In conclusion, MET and pMET (Y1003) were highly expressed in resected lung AC specimens. NS mutations were detected in all of the oncogenic genes assessed and the functional relevance of these mutations on tumorigenesis remains to be determined. Interestingly, novel MET mutations were detected in key functional domains; the SEMA domain which is critical for dimerization and ligand binding, and the JM domain which is important for MET downregulation via c-CBL. We will correlate these mutational and expression data with clinical outcomes in order to determine the prognostic role of MET.

Malignant pleural mesothelioma (MPM) is a devastating disease with an overall poor prognosis. Despite the recent advances in targeted molecular therapies, there is a clear and urgent need for the identification of novel mesothelioma targets for the development of highly efficacious therapeutics.In this study, we report that the expression of Sphingosine Kinase 1 (SphK1) protein was preferentially elevated in MPM tumor tissues (49 epithelioid and 13 sarcomatoid) compared to normal tissue (n = 13). In addition, we also observed significantly elevated levels of SphK1 and SphK2 mRNA and SphK1 protein expression in MPM cell lines such as H2691, H513 and H2461 compared to the non-malignant mesothelial Met5 cells. The underlying mechanism appears to be mediated by SphK1 induced upregulation of select gene transcription programs such as that of CBP/p300 and PCAF, two histone acetyl transferases (HAT), and the down regulation of cell cycle dependent kinase inhibitor genes such as p27Kip1 and p21Cip1. In addition, using immunoprecipitates of anti-acetylated histone antibody from SphK inhibitor, SphK-I2 treated Met5A and H2691 cell lysates, we also showed activation of other cell proliferation related genes, such as Top2A (DNA replication), AKB (chromosome remodeling and mitotic spindle formation), and suppression of p21 CIP1 and p27KIP1. The CDK2, HAT1 and MYST2 were, however, unaffected in the above study. Using SphK inhibitor and specific siRNA targeting either SphK1 or SphK2, we also unequivocally established that SphK1, but not SphK2, promotes H2691 mesothelioma cell proliferation. Using a multi-walled carbon nanotubes induced peritoneal mesothelioma mouse model, we showed that the SphK1-/- null mice exhibited significantly less inflammation and granulamatous nodules compared to their wild type counterparts.The lipid kinase SphK1 plays a positive and essential role in the growth and development of malignant mesothelioma and is therefore a likely therapeutic target.

Objective An area of need in cancer informatics is the ability to store images in a comprehensive database as part of translational cancer research. To meet this need, we have implemented a novel tandem database infrastructure that facilitates image storage and utilisation. Background We had previously implemented the Thoracic Oncology Program Database Project (TOPDP) database for our translational cancer research needs. While useful for many research endeavours, it is unable to store images, hence our need to implement an imaging database which could communicate easily with the TOPDP database. Methods The Thoracic Oncology Research Program (TORP) imaging database was designed using the Research Electronic Data Capture (REDCap) platform, which was developed by Vanderbilt University. To demonstrate proof of principle and evaluate utility, we performed a retrospective investigation into tumour response for malignant pleural mesothelioma (MPM) patients treated at the University of Chicago Medical Center with either of two analogous chemotherapy regimens and consented to at least one of two UCMC IRB protocols, 9571 and 13473A. Results A cohort of 22 MPM patients was identified using clinical data in the TOPDP database. After measurements were acquired, two representative CT images and 0–35 histological images per patient were successfully stored in the TORP database, along with clinical and demographic data. Discussion We implemented the TORP imaging database to be used in conjunction with our comprehensive TOPDP database. While it requires an additional effort to use two databases, our database infrastructure facilitates more comprehensive translational research. Conclusions The investigation described herein demonstrates the successful implementation of this novel tandem imaging database infrastructure, as well as the potential utility of investigations enabled by it. The data model presented here can be utilised as the basis for further development of other larger, more streamlined databases in the future.