Background: Lung cancer is the number one cancer killer worldwide. A major drawback in the lung cancer treatment field is the lack of realistic mouse models that replicate the complexity of human malignancy and immune contexture within the tumor microenvironment. Such models are urgently needed. Mutations of the tumor protein p53 are among the most common alterations in human lung cancers. Methods: Previously, we developed a line of lung cancer mouse model where mutant human TP53-273H is expressed in a lung specific manner in FVB/N background. To investigate whether the human TP53 mutant has a similar oncogenic potential when it is expressed in another strain of mouse, we crossed the FVB/N-SPC-TP53-273H mice to A/J strain and created A/J-SPC-TP53-273H transgenic mice. We then compared lung tumor formation between A/J-SPC-TP53-273H and FVB/N-SPC-TP53-273H. Results: We found the TP53-273H mutant gene has a similar oncogenic potential in lung tumor formation in both mice strains, although A/J strain mice have been found to be a highly susceptible strain in terms of carcinogeninduced lung cancer. Both transgenic lines survived more than 18 months and developed age related lung adenocarcinomas. With micro CT imaging, we found the FVB-SPC-TP53-273H mice survived more than 8 weeks after initial detection of lung cancer, providing a sufficient window for evaluating new anti-cancer agents. Conclusions: Oncogenic potential of the most common genetic mutation, TP53-273H, in human lung cancer is unique when it is expressed in different strains of mice. Our mouse models are useful tools for testing novel immune checkpoint inhibitors or other therapeutic strategies in the treatment of lung cancer.

ObjectiveWe aimed to evaluate and compare the impact of COVID-19 lockdown on lifestyle changes and other common related effects of the lockdown in Saudi adults with diabetes mellitus (DM), both type 1 (T1D) and type 2 diabetes (T2D).Methods265 T1D and 285 T2D individuals were included in this cross-sectional survey during lockdown using an online questionnaire and compared with 297 participants without DM. Variables included demographics, treatment changes, use of supplements, change in sleeping habits and physical activity, dietary changes, social and mental health, and education and awareness during COVID-19 lockdown.ResultsThe COVID-19 lockdown was associated with more treatment doses in people with T1D but not in those with T2D (p = 0.003). More participants with T1D and T2D than the control group reported that they felt symptoms of depression during lockdown (ORs of 1.83, p = 0.008 and 2.2, p = 0.001, respectively) and that lockdown affected them psychologically (ORs of 1.64, p = 0.019 and 1.85, p = 0.005, respectively). More participants with T1D than controls reported that their physical activity decreased during lockdown (OR of 2.70, p = 0.024). Furthermore, significantly lesser participants in both DM groups than controls agreed that the health education regarding COVID-19 covered everything (ORs of 0.41, p < 0.001 and 0.56, p < 0.001, respectively for T1D and T2D groups). Regarding dietary habits, the DM groups reported more changes in either the number of daily meals, meal content, or mealtimes than the control group.ConclusionsCOVID-19 lockdown-associated lifestyle changes were more prevalent in individuals with T1D and T2D compared to control. Findings may assist public health authorities in outlining their responses in pandemics and promote healthy lifestyle adaptations in this high-risk cohort to limit adverse effects in future lockdowns.

The parathyroid hormone 1 receptor (PTHR1) plays a crucial role in calcium homeostasis and bone metabolism. However, its genetic role in regulating bone turnover markers (BTMs) in postmenopausal osteoporosis (PMO) remains unclear. Herein, we explored parathyroid hormone (PTH) and PTHR gene variant susceptibility to osteoporosis and their association with various circulating BTM and inflammatory markers in postmenopausal women of Arab ethnicity. In total, 600 postmenopausal Arab women (300-PMO and 300-control) were genotyped for selected SNPs in PTH (rs1459015, rs307253, rs6054, rs307247, rs10500783 and rs10500784), PTHR1 (rs6442037, rs1138518, and rs724449 SNPs) and PTHR2 (rs9288393, rs10497900, and rs897083). Anthropometrics, BTMs, and inflammatory markers were measured. Bone mineral density (BMD) was measured at the lumbar spine L1–L4 and the femoral neck using dual-energy X-ray absorptiometry (DXA). PTHR1 rs1138518 genotype C/T was found to be a significant risk factor for PMO ( OR = 1.49 , 95% CI 1.0-2.1, P = 0.03 ). The genotypes C/T and T/T of PTHR1 rs1138518 were associated with 25-hydroxy-vitamin D (25(OH)D) regulation. In the PMO group, carriers of the C/T genotype had significantly lower 25(OH)D levels than carriers of the same genotypes in the control group (59.9 (36.7-92.4) nmol/l and 66.4 (43.5-87.8) nmol/l, respectively; P = 0.048 ]. Our study concludes that the PTHR1 rs1138518 genotype could be a potential risk factor for osteoporosis and 25(OH)D regulation in Arab women with PMO.

Introduction: A key response mechanism to DNA damage is the Fanconi Anemia repair pathway (FA), which involves homologous recombination DNA repair and is activated through mono- ubiquitination of FANCD2. FA deficiency is considered to increase the sensitivity of tumors to particular DNA-targeted agents, and may prove to be a target of cancer treatment. We hypothesize that FA deficient tumors have a low growth rate and reduced ability for DNA repair compared to FA functioning tumors. Given that genetic modifications can interfere with FA functionality, we aim to explore the association between the FA pathways and downstream genes that influence tumor growth. To date, few studies have examined gene expression associated with FA deficiency in lung cancer cells. Identification of the FA downstream genes may provide insight on DNA repair networks that impact cancer treatment. Methods: To generate FANCD2 knockdown cells, human lung cancer cell lines A549 and H1299 were transduced with FANCD2-specific short hairpin RNA expressing and puromycin-resistant lentiviral particles or control shRNA lentiviral particles. The cells were cultured in growth medium, and successful FANCD2 knockdown was confirmed by western immunoblot analysis. RNA deep sequencing was completed with Illumina RNA-Seq. We compared gene expression between knockdown FANCD2 and control samples across three cell lines and ranked significant gene expression changes, defined as a five-fold change in upregulation or downregulation. The fold change was calculated by dividing FANCD2 deficient expression by FANCD2 efficient expression. Results and discussion: 13436 genes were evaluated across three cell lines and 17 genes demonstrated gene expression change by at least 5-fold with FANCD2 knockdown in three cell lines. FANCD2 knockdown resulted in 14 downregulated genes and 3 upregulated genes. The downregulated genes RP11-618G20.1, RP5-1021I20.4, RP11-219A15.1, XXbac-BPG32J3.20, and BMS1P17 demonstrated significant expression change across three cell lines. Of the 14 downregulated genes, 13 genes had literature supporting oncogenic function. FA downstream genes confers oncogenic function. As FANCD2 is considered to promote cell proliferation, downregulation of oncogenic genes expression was expected with FANCD2 knockdown. However, the literature suggested that the 3 upregulated genes with FANCD2 knockdown also have oncogenic function. These genes may have functioning beyond the scope of carcinogenesis which may explain gene upregulation with FANCD2 knockdown. Pinpointing genes related to FA pathway deficiency may provide insight into genetic phenomena that drive cancer. Our results provide a starting point for developing targets to specific downstream genes associated with FA deficient tumors, which may prove to limit cancer progression. Further investigation is needed to determine how FANCD2 interacts with these genes to promote cell proliferation. Citation Format: Bianca Nguyen, Li Gao, Abeer Almiman, Shirley Tang, Kathleen Dotts, Miguel A. Villalona-Calero, Wenrui Duan. Investigation of Fanconi Anemia pathway downstream genes [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2568.

Background: Lung cancer is the number one cancer killer worldwide. A major impediment to progress in the lung cancer treatment field is the lack of realistic mouse models that replicate the complexity of human malignancy and immune contexture within the tumor microenvironment. Such models are urgently needed. Mutations of the tumor suppressor gene TP53 are among the most common alterations in human lung cancers. Methods: Previously, we developed a line of lung cancer mouse model where mutant human TP53-273H is expressed in a lung specific manner in FVB/N background. To investigate whether the human TP53 mutant has a similar oncogenic potential when it is expressed in another strain of mouse, we crossed the FVB/N-SPC-TP53-273H mice to A/J strain and created A/J-SPC-TP53-273H transgenic mice. We then compared lung tumor formation between A/J-SPC-TP53-273H and FVB/N-SPC-TP53-273H. Results: We found the TP53-273H mutant gene has a similar oncogenic potential in lung tumor formation in both mice strains, although A/J strain mice have been found to be a highly susceptible strain in terms of carcinogen-induced lung cancer. Both transgenic lines survived more than 18 months and developed age related lung adenocarcinomas. With micro CT imaging, we found the FVB-SPC-TP53-273H mice survived more than 8 weeks after initial detection of lung cancer, providing a sufficient window for evaluating new anti-cancer agents. Conclusions: Oncogenic potential of the most common genetic mutation, TP53-273H, in human lung cancer is unique when it is expressed in different strains of mice. Our mouse models are useful tools for testing novel immune check point inhibitors or other therapeutic strategies in treatment of lung cancer.

Introduction: The pseudokinase Tribbles pseudokinase 3 (TRIB3) is known as a regulator in cellular responses to a variety of stresses such as glucose insufficiency and (ER) stress. TRIB3 has been described in some studies as a tumor suppressor due to its effect on inactivating PI3K/Akt pathway, but other studies suggest TRIB3 as a stimulator of tumor progression. In this study, we aimed to define the functions of TRIB3 in in non-small cell lung cancer. Methods: TRIB3 expression was altered using a lentiviral vector to overexpress TRIB3 in non-small cell lung cancer. A CRISPR-CAS9 construct with guiding sequence matching to TRIB3 gene was used to knock out its expression. Cell proliferation was evaluated using MTS and trypan blue assays. Boyden chamber assay was used to assess the cell migration while cell cycle phases were determined using flow cytometry. Heatmap analysis was performed to assess the changes in the expressions of genes in cell cycle progression and apoptosis. Results: TRIB3 overexpression led to reduced cell proliferation and migration. Moreover, it contributed to the increased of cell cycle arrest at G0/G1 phase. QPCR analyses of cell cycle related genes showed an upregulation of CDK inhibitors in (NCI-H358) TRIB3 overexpression cells, while depletion of TRIB3 led to the down-regulation of CDK inhibitors. TRIB3 overexpression led to downregulation of LC3B and other autophagy markers while increasing the expression of apoptotic genes. Conclusion: Increased TRIB3 expression in non-small cell lung cancer cells inhibited proliferation by blocking cell cycle progression through the upregulation of CDK inhibitors, led to activation of cell death through apoptosis. Our study reveals a significant role of TRIB3 in regulating cell cycle progression, apoptosis and autophagy. Citation Format: Abeer Almiman, Daotai Nie, Jamila Adom. TRIB3 regulates cell cycle progression and programmed cell death in non-small cell lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 307. doi:10.1158/1538-7445.AM2017-307

In 2018, 154,100 lung cancer deaths are projected to occur in the United States. Currently, there are very few predictors to identify lung cancer in its early stages. Recent studies have demonstrated a link between microRNAs (miRNAs) and specific cancers. Dysregulation of miRNAs in cancer can lead to upregulate oncogenes and downregulate tumor suppressors. Previously, we developed a line of transgenic mice expressing the human mutant p53 (273H) gene. The transgenic mice were observed to have accelerated risk of spontaneous adenocarcinomas at the age of 13-14 months. However the mechanisms involved in lung tumorigenesis at this age cohort is still unclear. Herein, we report our analysis on miRNA expression in lung tissue between a low risk cohort and high risk cohort. To investigate the role of mutant p53 in lung tumorigenesis, a p53(273H) transgenic mouse model was developed. Murine lung tissue were harvested from age groups of 7 months (low risk group with risk of 5% of lung cancer), and the 13.5 months (high risk group with risk of 27% of lung cancer). RNA was then extracted with trizol, and miRNAs were quantified using a Nanostring counter miRNA array. The miRNA expressions were normalized and averaged. The normalized miRNAs from high risk group (n=3) were compared with the miRNAs obtained from the low risk group (n=3). We screened 600 different miRNA expressions from these age cohorts. Total of 19 miRNAs that had an up-regulation of at least 5-fold in the high risk cohort as compared with the low risk cohort including some important miRNAs in human lung cancer (e.g mmu-miR-2133, mmu-miR-2140, mmu-miR-142-5p, mmu-miR-2138, mmu-miR-2134, mmu-miR-2135, mmu-miR-452, mmu-miR-703, mmu-miR-690, mmu-miR-206, mmu-miR-706, mmu-miR-691, mmu-miR-208a, mmu-miR-574-5p, mmu-miR-1). Four miRNAs had a down regulation at least 5-fold in the high risk cohort including mmu-miR-720, mmu-miR-1937c, and mmu-miR-1937a+mmu-miR-1937b.We found a group of microRNAs to be expressed abnormally high and another group microRNA to be expressed abnormally low in the high risk group as comparing to the low risk group. We compared the data to our murine lung cancer microRNA profile and found a group of miRNA expression potentially correlated to risk of lung cancer development in mice. Most human lung cancers are at an advanced stage when they are first found. These cancers are very hard to cure. However, if a lung cancer is found at an earlier stage when it is small and before it has spread, the early stage lung cancer can be successfully treated. Further studies are necessary to employ these miRNAs as early diagnostic biomarkers in prediction of human lung cancers. Citation Format: Myia Aiges, Abeer Almiman, Li Gao, Kathleen Dotts, Miguel A. Villalona-Calero, Wenrui Duan. Differential miRNA expression levels and risk of lung adenocarcinoma in transgenic mice [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 3548.

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