Lung cancer currently has the greatest mortality rate of cancer patients of all sexes in the United States (Torre et al., 2016). Low-dose CT scans are utilized for lung cancer screening in patients who fall within the NLST entry criteria (Sanchez-Salcedo et al., 2015). The original criteria for screening were age over 55 and pack-year over 30, which were recently changed to age 50 and pack-year over 20 in 2021. The study population in this paper utilized the original criteria. A system developed and copyrighted by the American College of Radiology (ACR) referred to as the Lung CT Screening Reporting and Data System (Lung-RADS) has implemented a standardized method of classifying and interpreting lung cancer chest CT screening results. Lung-RADS has a scoring system which is scaled 1 – 4x (Pinsky et al., 2015) The likelihood of malignancy based on nodule appearance, diameter, and presence of growth comprise the components of which score is given (Chung et al., 2017). Lung-RADS 2 scored nodules are benign nodules and patients follow up for another CT in a year. Lung-RADS 3 nodules are probably benign nodules; however, they do have a low-risk of malignancy. It is known that a select few of these relatively benign appearing nodules will turn out to be malignant. Lung-RADS 4 nodules have a >5% chance of malignancy and can be confirmed through pathology. In this project, a retrospective chart review analyzing patient demographics and pulmonary health history will be correlated to lung-RADS risk likelihood of malignancy. Machine learning will also be utilized to study and analyze radiographic factors associated with the sample. The CT scans of patients who previously scored in an intermediate category will be compiled and analyzed to determine potential common demographical, clinical and radiomic factors which will hopefully allow intermediately categorized nodule indicators to be used to detect cancers earlier and to more accurately classify lesions into benign or malignant categories. In all, the goal of this research is to determine common clinical, demographic and radiomic factors of patients who were deemed intermediate risk and then progressed to a higher categorization. The importance of expanding current risk factors for discrimination of benign from malignant will also be analyzed, along with those specific risk-factors within Lung-RADS intermediately categorized nodules. The characteristics and baseline co-morbidities of RADS 2 and 3 lung cancer patients by follow-up CT results, progression to RADS-4 on follow-up CT and lung cancer diagnosis will be compiled and exemplified.

<p>Cancer patients undergoing chemotherapy have an elevated risk for thrombosis. Although thrombosis is a common complication in cancer patients, the mechanisms of chemotherapy-induced thrombosis remain unclear. We investigated the procoagulant effects of lung cancer chemotherapy agents (carboplatin, paclitaxel, cisplatin, and gemcitabine) on endothelial cells, A549 cells, and monocytes. We also investigated the <em>in </em>vivo procoagulant effects of the aforementioned chemotherapeutic agents as well as the anti-angiogenic agent bevacizumab. Tissue factor (TF) activity, TF antigen and phosphatidylserine (PS) levels were measured on chemotherapy-treated human umbilical vein endothelial cells (HUVEC), A549 cells, and monocytes. Treatment of HUVECs, A549 cells, and monocytes with lung cancer single agent and combination chemotherapy resulted in significant increases in TF activity. However, only cisplatin- and gemcitabine- treated monocytes were found to have increased TF antigen levels. PS exposure was increased only on HUVEC and monocytes treated with cisplatin/gemcitabine combination therapy. Interestingly, addition of paclitaxel to carboplatin resulted in reduced levels of PS exposure on monocytes. This study is the first to explore the procoagulant effects of lung cancer chemotherapy agents on monocyte and A549 cell TF activity levels, as well as to investigate the mechanisms by which lung cancer agents may promote TF decryption on these cell lines<strong>.</strong> Our <em>in vivo</em> results demonstrated that treatment of healthy mice with bevacizumab, paclitaxel and carboplatin moderately increased plasma TAT levels in healthy mice. These studies reveal potential mechanisms by which lung cancer chemotherapy may increase the risk of thrombosis. These studies reveal potential mechanisms by which lung cancer chemotherapy agents induce a hypercoagulable state.</p> / Master of Science (MSc)

<p>Brain metastases are most common in adults suffering from lung cancer, predicting uniformly poor patient outcome and short survival time. Despite their frequency and severity, very little is known about the tumorigenesis of brain metastases. Previously developed primary brain tumour-initiating cell (BTIC) models were used to determine the presence of a stem-like population in brain metastases from the lung. Use of clinical samples and the NCI-H1915 cell line allowed for the development of useful strategies for study of brain metastasis.</p> <p>The sphere formation capacity and expression of known BTIC markers in brain metastases was suggestive of a self-renewing population. Differentiation studies demonstrated that neither clinical samples nor NCI-H1915 cells had neural lineage potential. Intracranial xenotransplant of clinical samples and NCI-H1915 cells into NOD-SCID mice led to formation of multiple focal masses throughout the ventricles; the tumours were also serially transplantable, further implicating a TIC population. Of known BTIC markers, only CD15 expression levels and patterns were similar enough in clinical samples and NCI-H1915 cells to warrant prospective sorting experiments in the cell line. Use of CD15 failed to identify a CSC or TIC population in NCI-H1915 cells.</p> <p>These findings suggest that a TIC population is present in brain metastases; however, this remains to be identified. It is recommended that due to the limitations of cell surface markers, the study of brain metastasis should use a selective gene expression approach, in order to target genes and pathways essential to metastasis. It was shown that NCI-H1915 cells could be useful for such an approach, studying the effects on proliferation, sphere formation, and tumour formation capacity of brain metastases from the lung. Further study using this model could ultimately lead to the disruption of pathways essential to the metastatic process, transforming a uniformly fatal disease into a more localized and treatable one.</p> / Master of Science (MSc)

<p><strong><em>Purpose</em></strong></p> <p>To examine the potential of the anti-diabetic agent Metformin (MET) to enhance responses of NSCLC to ionizing radiation (IR).</p> <p><strong><em>Experimental Design</em></strong></p> <p>Human NSCLC A549, H1299 and SK-MES cells were treated with IR, MET or the mTOR inhibitor rapamycin and subjected to proliferation, clonogenic, immunoblotting, cell cycle and apoptosis assays. A549 and H1299 cells were grafted into flanks of immunosuppressed mice and treated with MET and/or IR. Tumours were analyzed by immunoblotting and immunohistochemistry.</p> <p><strong><em>Results</em></strong></p> <p>MET(2.5uM-5mM) caused dose-dependent inhibition of proliferation (10-70%)in all lines, inibited clonogenic survival and sensitized cells to IR. In A549 cellsMET caused inhibition of proliferation comparable to rapamycin, stimulated expression and activation of the ATM and AMPK-p53-p21^cip1and inhibited the Akt-mTOR-4-EBP1 pathway.MET caused G1 arrest of cell cycle, enhanced apoptosis and induced sustained DNA repair foci of gH2AX. MET and IR alone inhibited xenograft growth and combined treatment enhanced that further. IR and MET induced sustained enhancement of expression and activity of ATM-AMPK-p53-p21^cip1and inhibitionof Akt-mTOR-4-EBP1 pathways in tumours also. MET reduced expression of angiogenesis and enhanced expression of apoptosis markers in both control and radiated tumours.</p> <p><strong><em>Conclusions</em></strong></p> <p>Clinically achievable(uM) doses ofMET inhibit human NSCLC cell and tumour growth and sensitize them to IR.This is accompanied by desirable modulation of molecular signals, inhibition of angiogenesis and induction of apoptosis. Our results suggest that MET could be a clinically useful adjunct to radiotherapy in NSCLC and support clinical investigation of MET in combination with radiotherapy.</p> / Master of Science (MSc)

<p> It has been suggested that combination treatment of high dose rate (HDR) intraluminal brachytherapy and PDT (Photodynamic therapy) in non-small cell lung cancer (NSCLC) may improve the efficacy of treatment, reduce the toxicity and improve quality of life for patients. To provide a cellular basis for this approach we have examined the in vitro sensitivity of normal lung fibroblasts (MRC5) and four NSCLC cell lines (SKMES-1, A549, NCIH460 and NCIH23) following, UVC treatment, HDR radiation, HDR radiation with Photofrin alone, PDT and combined HDR radiation and PDT. Cell sensitivity was measured using clonogenic survival. HDR radiation was cobalt-60 gamma rays (1.5-1.9 Gy/min). For PDT treatment, cells were exposed to 2.5 J.lg/ml Photofrin for 18-24 h followed by light exposure (20mW/cm2). D37 values calculated from the survival curves indicated a 2-fold difference in sensitivity to UVC, 6-fold difference in HDR radiation sensitivity and an 8-fold difference in PDT sensitivity. All cell lines showed a similar Photofrin uptake per cell when measured by flow cytometry using 488nm excitation and 620-675 nm emission wavelengths. Photofrin alone at concentrations up to 10 J.lg/ml had no significant effect on the survival of the NSCLC cell lines, whereas 10 J.lg/ml ofPhotofrin alone reduced survival significantly in MRC5 cells. A radiosensitizing effect of Photofrin was detected in MRC5 and NCIH460 cells, but not in A549, SKMES-1 and NCI-H23 cells. For combined treatment cells were exposed to Photofrin and then either exposed to light and 15-30 minutes later exposed to HDR radiation or exposed to HDR radiation and 15-30 minutes later exposed to light. Results showed that although light followed by gamma rays resulted in a somewhat greater tumor cell kill compared to gamma rays followed by light this difference was not significant for any of the cell lines tested. However, this difference was significant when data for all NSCLC cell lines were pooled. The combined treatment with high dose rate HDR radiation and PDT was not significantly different from an additive effect of the individual treatment modalities for in vitro survival of 4 NSCLC cells. In contrast the combined treatment was less than additive for the MRCS cells suggesting that the combined treatment would have the potential advantage of doing less damage to the normal lung cells and suggests that equivalent tumour cell kill in vivo may be possible at reduced systemic effects to patients. In preliminary experiments we have started to examine the effects of Photofrin-mediated PDT on the extra cellular signal-activated protein kinase (ERK) signaling pathway in NSCLC cells. The use of multiple NSCLC cell lines allows for the possible identification of cell line specific changes involved in resistance to PDT and HDR radiation and this will be explored in future work. </p> / Thesis / Master of Science (MSc)

No description available.

Cancer is one of the main causes of death in the US. Lung cancer remains the highest killing form of cancer. Lung screening rates are low amongst the general population and even lower in minority populations. It is not well known what the barriers are for lung cancer screening. In order to investigate barriers, I created questions to add onto an existing questionnaire survey for an ongoing lung cancer screening research project. These questions focused on social determinants of health and the survey was administered to patients who were non-adherent to lung cancer screening. Patients reported cost concerns for screening, potential medical care costs, and ability to attend medical appointments as barriers to lung cancer screening. Both non-white and female patients reported more difficulties attending appointments than their white and male counterparts. Patient physician relationship and perceived racial discrimination were not barriers to lung cancer screening in the patient population surveyed. Further research needs to investigate specific details on these barriers to create interventions to increase lung cancer screening rates. / Urban Bioethics

Since the dawn of the genomics era, the genetics of numerous human disorders has been understood which has led to improvements in targeted therapeutics. However, the focus of most research has been primarily on protein coding genes, which account for only 2% of the entire genome, leaving much of the remaining genome relatively unstudied. In particular, repetitive sequences, called microsatellites (MST), which are tandem repeats of 1 to 6 bases, are known to be mutational hotspots and have been linked to diseases, such as Huntington disease and Fragile X syndrome. This work represents a significant effort towards closing this knowledge gap. Specifically, we developed a next generation sequencing based enrichment method along with the supporting computational pipeline for detecting novel MST sequences in the human genome. Using this global MST enrichment protocol, we have identified 790 novel sequences. Analysis of these novel sequences has identified previously unknown functional elements, demonstrating its potential for aiding in the completion of the euchromatic DNA. We also developed a disease risk diagnostic using a novel target specific enrichment method that produces high resolution MST sequencing data that has the potential to validate, for the first time, the link between MST genotype variation and cancer. Combined with publicly available exome datasets of non-small cell lung cancer and 1000 genomes project, the target specific MST enrichment method uncovered a signature set of 21 MST loci that can differentiate between lung cancer and non-cancer control samples with a sensitivity ratio of 0.93. Finally, to understand the molecular causes of MST instability, we analyzed genomic variants and gene expression data for an autosomal recessive disorder, Fanconi anemia (FA). This first of its kind study quantified the heterogeneity of FA cells and demonstrated the possibility of utilizing the DNA crosslink repair dysfunctional FA cells as a suitable system to further study the causes of MST instability. / Ph. D.

<p dir="ltr">Coronavirus disease 2019 (COVID-19) is a serious public health threat, and lung cancer patients are at high risk of poor outcomes from COVID-19 relative to the general population. Little is known about lung cancer patients’ beliefs and emotions regarding COVID-19 and COVID-19 protective behaviors (i.e., mask wearing, social distancing, hand hygiene). Prior research has found that Health Belief Model (HBM) variables (i.e., perceived risk of acquiring the illness, perceived illness severity, perceived benefits of and barriers to the preventive health behavior) and worry are predictive of engagement in preventive health behaviors. Drawing upon the HBM and theories of the role of emotion in decision-making, the current study examined psychological correlates of lung cancer patients’ engagement in COVID-19 protective behaviors. Lung cancer patients (<i>N</i> = 191) were recruited from Indiana University Simon Comprehensive Cancer Center to participate in a one-time survey from August 2021 through May 2022. Descriptive statistics were used to characterize COVID-19 protective behaviors. Multiple hierarchical regression models were used to examine associations between HBM constructs and engagement in COVID-19 protective behaviors. Higher-order regression models were then used to examine whether worry about COVID-19 was associated with COVID-19 protective behaviors above and beyond the effects of HBM variables. In general, the present sample showed high rates of engagement in all COVID-19 protective behaviors. Fewer perceived barriers to mask wearing were associated with greater mask wearing, and greater worry about COVID-19 was associated with greater mask wearing above and beyond the effects of HBM variables. Greater perceived severity of COVID-19 was associated with more social distancing, and higher levels of worry about COVID-19 were associated with more social distancing above and beyond the effects of HBM variables. HBM constructs (i.e., perceived risk of COVID-19, perceived severity of COVID-19) and worry about COVID-19 were unrelated to hand hygiene. Most findings are in line with theories on the central role of emotion in health-related decision-making and warrant replication in longitudinal research. Results point to several potential intervention targets, including worry about COVID-19, perceived severity of COVID-19, and perceived barriers to mask wearing, to improve COVID-19 protective behaviors in lung cancer patients..<br></p>

No / Improved understanding of the involvement of matrix metalloproteinases (MMPs), including membrane-type MMPs (MT-MMPs), in human tumours has potential diagnostic, prognostic and therapeutic implications. We assessed the relationship between MT-MMP expression and clinicopathological parameters in human non-small cell lung cancer (NSCLC) and histologically normal lung tissue by quantitative Real Time PCR (qRT-PCR). All MT-MMPs (MMPs 14-17, 24 and 25) were detected by qRT-PCR with significantly higher MMP-14, -15 and -17 expression observed in tumour relative to normal lung specimens. MMP-16 was undetectable in normal lung but expressed in 8% tumours. MMP-15 demonstrated significant overexpression in adenocarcinomas relative to squamous cell carcinomas and normal lung tissue. MMP-14 mRNA expression strongly correlated to MMP-14 proteolytic activity in preclinical tumour models, indicating that qRT-PCR may predict MMP-14 activity levels in NSCLC. These data suggest that MMP-14, -15 and -17 may be good markers of disease, or therapeutic targets for treatment of human NSCLC.