Non-small Cell Lung Adenocarcinoma Research Articles

Human epididymis protein 4 may not be a reliable screening biomarker for detecting lung carcinoma patients.

Human epididymis protein 4 (HE4) acts as a protease inhibitor. It has been detected in the serum of patients with lung cancer patients and its utility for lung cancer screening was found in different studies. Nevertheless, little is known regarding the expression of HE4 in lung carcinoma subtypes. The aim of the present study was to investigate whether HE4 expression is a reliable marker for detecting any lung carcinoma subtypes, including small cell lung cancer (SCLC), non-small cell lung cancer (NSCLC) and adenocarcinoma (AC). In total, 141 lung carcinoma patients were enrolled in the study. Biopsy samples were obtained from bronchoscopic biopsy. The tumors were classified as SCLC (group 1, 54 cases) or NSCLC (group 2, 87 cases) based on histology and immunohistochemistry. The latter was sub-grouped as adenocarcinoma (group 2a, AC) and squamous cell carcinoma (group 2b, SCC). The immunohistochemical expression of HE4 was compared between the groups. The study revealed that the majority of the SCLC and SCC cases were devoid of HE4 (90.1 and 89.65%, respectively). Approximately 10% of cases had HE4 immune expression and the staining was focal and moderate throughout the tumor tissue. On the other hand, 78.8% of AC expressed HE4 and the staining was diffuse and strong. The overall HE4 expression in the lung cancer patients was 33.7%. In conclusion, the results of the present study have shown that HE4 is expressed in adenocarcinoma of the lung but it is not frequent in SCC and SCLC. The value of HE4 as a screening biomarker for lung cancer is limited but its presence exclusively in adenocarcinoma may provide new insight for targeted therapy.

Cytotoxic peptides with insulin-releasing activities from skin secretions of the Italian stream frog Rana italica (Ranidae).

Peptidomic analysis of norepinephrine-stimulated skin secretions from Italian stream frog Rana italica led to the purification and characterization of two host-defense peptides differing by a single amino acid residue belonging to the brevinin-1 family (brevinin-1ITa and -1ITb), a peptide belonging to the temporin family (temporin-ITa) and a component identified as prokineticin Bv8. The secretions contained relatively high concentrations of the methionine-sulphoxide forms of brevinin-1ITa and -1ITb suggesting that these peptides may have a role as antioxidants in the skin of this montane frog. Brevinin-1ITa (IVPFLLGMVPKLVCLITKKC) displayed potent cytotoxicity against non-small cell lung adenocarcinoma A549 cells (LC50 =18μM), breast adenocarcinoma MDA-MB-231 cells (LC50 =8μM) and colorectal adenocarcinoma HT-29 cells (LC50 =18μM), but the peptide was also strongly hemolytic against mouse erythrocytes (LC50 =7μM). Temporin-ITa (VFLGAIAQALTSLLGKL.NH2 ) was between three and fivefold less potent against these cells. Brevinin-1ITa inhibited growth of both Gram-positive Staphylococcus epidermidis and Gram-negative Escherichia coli as well as a strain of the opportunist yeast pathogen Candida parapsilosis, whereas temporin-ITa was active only against S. epidermidis and C. parapsilosis. Both peptides stimulated the release of insulin from BRIN-BD11 clonal β-cells at concentrations ≥1nM, but brevinin-1ITa was cytotoxic to the cells at concentrations ≥3μM. Copyright © 2017 European Peptide Society and John Wiley & Sons, Ltd.

Abstract 920: COP1 E3 ligase regulates response to oncogenic MAPK pathway inhibition

Abstract Oncogenically activated RAS-MAPK pathway is the driver of several cancers including the majority of non-small cell lung adenocarcinomas (NSCLC). RAS-MAPK pathway is activated in NSCLC tumors via diverse genetic alterations in upstream receptor tyrosine kinases such as EGFR and ALK as well as in RAS, BRAF, MEK and RAS GTPase activating protein (GAP) and tumor suppressor, NF1. Molecular targeting of players of RAS-MAPK pathway can elicit an initial tumor response in many patients. However, most patients show an incomplete response and some fail to respond despite the presence of RAS-MAPK pathway activating genetic lesion in the tumor. Hence, it is important to understand the molecular basis of response and resistance to targeted therapies inhibiting the RAS-MAPK pathway in lung adenocarcinomas in order to improve patient survival. We conducted a genetic screen to identify regulators of response to MAPK pathway inhibition in lung adenocarcinomas. Our genetic screen uncovered the E3 ubiquitin ligase COP1/RFWD2 as a novel modulator of response to inhibition of RAS-MAPK pathway. We found that depletion of COP1 and members of its complex, as well as proteasomal subunits, confers resistance to RAS-MAPK pathway inhibition in human lung adenocarcinoma cells with oncogenically activated RAS-MAPK pathway. Interestingly, oncogenic targets of COP1 include MAPK pathway effectors, presence of which has been shown to support the survival of cancer cells with oncogenically activated MAPK upon inhibition of the RAS-MAPK pathway. Hence, we tested if depletion of COP1 alters the levels of those oncogenic substrates. Excitingly, we observed a substantial impact of COP1 depletion on the levels of certain effectors in the presence of RAS-MAPK inhibitors in genetically diverse NSCLC cells and also in oncogenic-BRAF driven melanoma cells. Our studies suggests that depletion of COP1 confers resistance to MAPK pathway inhibition in RAS-MAPK pathway driven cancers through accumulation of specific MAPK pathway effectors. This work has improved our understanding of the molecular basis of tumor cell resilience during initial treatment as well as of primary treatment resistance. Additionally, we are examining if levels of COP1 could be a biomarker for predicting response to RAS-MAPK pathway inhibitor therapy. We will also determine if the resistance-conferring COP1 substrate could be cotargeted along with RAS-MAPK pathway to improve the patient response in RAS-MAPK pathway driven lung adenocarcinomas and other cancers with low levels of COP1. Citation Format: Manasi K. Mayekar, Luping Lin, Trever G. Bivona. COP1 E3 ligase regulates response to oncogenic MAPK pathway inhibition [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 920.

Abstract 3770: CAR-T cell harboring a camelid single domain antibody as a targeting agent to kill tumors expressing VEGFR2

Abstract Modulation of the immune system is showing tremendous promise in the treatment of malignancies. In addition to checkpoint inhibitors that re-activate T cells present in the tumor microenvironment, exogenously transduced chimeric antigen receptor (CAR) T cells are providing excellent responses in clinical trials for the treatment of leukemias. In this study, we describe CAR-T cells that target VEGFR2-expressing tumors. Angiogenesis is the process of new blood vessel formation and is essential for a tumor to grow beyond a certain size. Tumors secrete the pro-angiogenic factor vascular endothelial growth factor (VEGF), which acts upon local endothelial cells by binding to vascular endothelial growth factor receptors (VEGFR). As VEGFR2 is also expressed by a variety of tumors, we investigated the utility of anti-VEGFR2 CAR-T cells as a method to treat VEGFR2-expressing tumors. Camelid antibodies are small (14 kD) single chain antibodies. To generate a camelid antibody targeting the extracellular domain of VEGFR2, a llama was immunized with recombinant VEGFR2/Fc. A phage display library was generated and screened to identify an antibody with high binding affinity to VEGFR2. The selected antibody was expressed in the E. coli. BL21 (DE3) pT7 system. The purified antibody was characterized by SEC, LC-MS peptide mapping and ELISA. CAR-T cells were engineered to express the camelid anti-VEGFR2 antibody in combination with the CD28 and 4-1BB costimulatory molecules and the CD3 zeta chain. Tumor cells were screened for expression of VEGFR2, and the HL-60 acute promyelocytic leukemia, ZR-75-30 breast ductal carcinoma and NCI-H23 non-small cell lung adenocarcinoma were identified. Co-incubation of anti-VEGFR2 CAR-T cells with the VEGFR-2-expressing cell lines resulted in dose-dependent target cell toxicity as measured by LDH release. In addition, T cell activity was confirmed, as high levels of IL-2 and IFN-γ were detected in the cell culture media. These results suggest that anti-VEGFR2 CAR-T may be useful in directly targeting VEGFR2-expressing tumors. We previously showed the utility of camelid antibodies in CAR-T constructs as anti-CEACAM6 CAR-T cells show both in vitro and in vitro efficacy against the pancreatic tumor Bx-PC3. The use of the a camelid V21 antibody to target VEGFR2-expressing tumors provides hope that camelid single domain antibodies can be developed for CAR-T therapies. Citation Format: Heman Chao, Baomin Tian, Marni Uger, Wah Wong. CAR-T cell harboring a camelid single domain antibody as a targeting agent to kill tumors expressing VEGFR2 [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 3770. doi:10.1158/1538-7445.AM2017-3770

Associations between radiologist-defined semantic and automatically computed radiomic features in non-small cell lung cancer

Tumor phenotypes captured in computed tomography (CT) images can be described qualitatively and quantitatively using radiologist-defined “semantic” and computer-derived “radiomic” features, respectively. While both types of features have shown to be promising predictors of prognosis, the association between these groups of features remains unclear. We investigated the associations between semantic and radiomic features in CT images of 258 non-small cell lung adenocarcinomas. The tumor imaging phenotypes were described using 9 qualitative semantic features that were scored by radiologists, and 57 quantitative radiomic features that were automatically calculated using mathematical algorithms. Of the 9 semantic features, 3 were rated on a binary scale (cavitation, air bronchogram, and calcification) and 6 were rated on a categorical scale (texture, border definition, contour, lobulation, spiculation, and concavity). 32–41 radiomic features were associated with the binary semantic features (AUC = 0.56–0.76). The relationship between all radiomic features and the categorical semantic features ranged from weak to moderate (|Spearmen’s correlation| = 0.002–0.65). There are associations between semantic and radiomic features, however the associations were not strong despite being significant. Our results indicate that radiomic features may capture distinct tumor phenotypes that fail to be perceived by naked eye that semantic features do not describe and vice versa.

Inflamed actinic keratoses associated with pemetrexed and carboplatin therapy

Eruptive actinic keratosis (AK) consequent to systemic chemotherapy can be confused with drug allergies. We present the first case of inflamed AKs in one patient after receiving combination therapy with pemetrexed and carboplatin.A 68-year-old woman with non-small cell lung adenocarcinoma (NSCLC) presented with numerous pruritic ill-defined, gritty, erythematous papules consistent with AKs on her upper chest, upper back, and arms two weeks after completing the first cycle of combination therapy with carboplatin and pemetrexed. The care team managed her with topical steroids and the lesions resolved within one month. The patient resumed the second cycle of chemotherapy and reported the occurrence of a similar but milder eruption.This case illustrates that eruptive AKs should be considered in the differential diagnosis of drug-related rashes, especially if the physical exam is suggestive. The mainstay of treatment should be directed at symptomatic improvement, and chemotherapy may be continued.

Homeopathy: Curative, Concurrent and Supportive Cancer Treatment Potential

Background: Homeopathy is used by 12 to 24% of European cancer patients, representing 40.4% of patients at European integrative cancer centers. In 2011, a Swiss literature review on homeopathy led to homeopathic treatment coverage in the Swiss national health insurance program. Homeopathy for curative pediatric cancer treatment is limited to 7.4% in the Netherlands, but, 76.5% of German parents will use homeopathy as part of their children’s cancer treatment. The purpose of this paper is to determine what is needed for homeopathy to play a larger role in curative, concurrent, and supportive cancer treatment. Methods: PubMed searches in September 2016 and January 2017 were performed with search terms “adverse effects, breast cancer, cancer, cervical cancer, endometrial cancer, homeopathy, ovarian cancer, prevention, treatment”. Curative, concurrent, and supportive homeopathic cancer treatments material was taken from these searches. Findings: At least five homeopathic formulations are immunologic adjuvants, activating natural killer cell destruction of cancer and virally infected cells. Ultramolecular Carcinosin, Phytolacca decandra, Conium, Thuja and Klimaktoplan® are appropriate for in vivo breast cancer trials. Lycopodium clavatum 5C and 15C are ready for in vivo cervical cancer trials. Sulphur 30C, may be considered for non-small cell lung adenocarcinoma treatment trials. Conventional cancer treatment associated anxiety, asthenia, depression, dermatitis, folliculitis, hot flushes, insomnia, nausea and vomiting, and stomatitis, respond to numerous homeopathic treatments including hetero-isotherapy. Conclusion & Significance: In vitro studies and retrospective case series indicate that homeopathy could provide curative cancer treatment for an array of cancers: Breast, cervix, gallbladder, liver, lung, oral, pancreas, periampullary, skin, and stomach. Appropriately designed randomized controlled trials (RCT) based on reproducible homeopathic treatments and clinical protocols, with intent-to-treat analysis will have increased validity. If these RCT have positive outcomes homeopathy will secure a position in curative, concurrent, and supportive cancer treatment.

Anti-Proliferative and Apoptosis-Inducing Effect of Theabrownin against Non-small Cell Lung Adenocarcinoma A549 Cells.

With the highest cancer incidence rate, lung cancer, especially non-small cell lung cancer (NSCLC), is the leading cause of cancer death in the world. Tea (leaves of Camellia sinensis) has been widely used as a traditional beverage beneficial to human health, including anti-NSCLC activity. Theabrownin (TB) is one major kind of tea pigment responsible for the beneficial effects of tea liquor. However, its effect on NSCLC is unknown. The aim of the present study was to evaluate anti-proliferative and apoptosis-inducing effect of TB on NSCLC (A549) cells, using MTT assay, morphological observation (DAPI staining), in situ terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay, and annexin-V/PI flow cytometry. Subsequently, the expression of several genes associated with cell proliferation and apoptosis were detected by real time PCR assay to explore its potential underlying mechanism. TB was revealed to inhibit cell proliferation of A549 cells in a concentration-dependent and time-dependent manner. Morphological observation, TUNEL assay and flow cytometric analysis evidenced an apoptosis-inducing effect of TB on A549 cells in a concentration-dependent manner. The real time PCR assay demonstrated that TB down-regulated the expression of TOPO I, TOPO II, and BCL-2, and up-regulated the expression of E2F1, P53, GADD45, BAX, BIM, and CASP 3,7,8,9, which suggests an activation of P53-mediated apoptotic (caspase-dependent) pathway in response to TB treatment. The western blot analysis showed a similar trend for the corresponding protein expression (P53, Bax, Bcl-2, caspase 3,9, and PARP) and further revealed DNA damage as a trigger of the apoptosis (phosphorylation of histone H2A.X). Accordingly, TB can be speculated as a DNA damage inducer and topoisomerase (Topo I and Topo II) inhibitor that can up-regulate P53 expression and subsequently modulate the expression of the downstream genes to induce cell proliferation inhibition and apoptosis of A549 cells. Our results indicate that TB exhibits its anti-NSCLC activity via a P53-dependent mechanism, which may be a promising candidate of natural product for anti-cancer drug development in the treatment of NSCLC.

Choroidal metastasis as a presenting manifestation of a lung adenocarcinoma with response to afatinib

Abstract Case report We present the case of a 55-year-old man with a non-small cell lung adenocarcinoma, who presented with choroidal metastasis. The patient showed a decrease in visual acuity. His evaluation revealed unilateral choroidal metastasis secondary to carcinoma of the lung. The patient received afatinib with complete regression of choroidal metastasis after one year follow-up. Discussion Choroidal metastasis may be the initial sign of lung cancer. This case highlights the importance of a thorough systemic evaluation in patients with choroidal tumors. Afatinib was effective against choroidal metastasis of a lung adenocarcinoma with EFGR mutation.

Abstract 3498: Increased cellular stress and apoptosis induced by anti-SIAH2 therapy in human cancer cells with oncogenic K-RAS addiction

Abstract Hyperactivated K-RAS oncoproteins are known to drive cell proliferation, tumor growth, and metastasis. Oncogene addiction is often associated with increased ER and mitochondrial stress due to excessive reactive oxygen and nitrogen species (ROS & NROS) production. RING finger E3 ubiquitin ligase, Seven in Absentia homolog 2 (Siah2) is a highly conserved, critical signaling module downstream of the EGFR/HER2/K-RAS pathway. SIAH2 function is required for proper K-RAS signal transduction and has been shown to target key regulators of oxidative stress for ubiquitination and proteasome degradation. However, the mechanisms by which SIAH2 regulates cellular stress and protects against programmed cell death in oncogenic K-RAS-addicted cancer cells are not well understood. Since SIAH2 is upregulated in proliferating tumor cells, we hypothesized that SIAH2-deficient cancer cells may be compromised in their ability to adapt to increased cellular and oxidative stress induced by oncogenic K-RAS pathway blockade. In this study, SIAH2 knockdown was carried out in cancer cells carrying oncogenic K-RAS mutations (A549, MDA-MB-231 and MiaPaCa cells). Triplicates of fresh cell lysates from SIAH2 proficient and SIAH2 deficient cells were then interrogated for altered expression of 19 signaling molecules with known functions in modulating stress response and apoptosis using the PathScan® Stress and Apoptosis Signaling Antibody Array kit. Differences in the protein expression were quantified and validated by Western blots, immunofluorescence (IF) and flow cytometry. We found that oxidative stress was markedly increased, evidenced by mitochondrial aggregations and dysfunction in a subset K-RAS-driven cancer cells, as we blocked SIAH2 function downstream of the oncogenic K-RAS signaling pathway. SIAH2 deficiency in breast cancer (MDA-MB-231) and non-small cell lung adenocarcinoma (A549) cells resulted in decreased ERK1/2 and AKT (S473) phosphorylation, increased ROS production, compromised mitochondria integrity and function, increased Caspase 3 and 7 activity, and PARP 1 cleavage that led to massive cell death observed in these cancer cells. In contrast, the prominent cell death was not observed in SIAH2 deficient pancreatic cancer (MiaPaCa) cells that expressed high levels of Survivin, which is an anti-apoptosis protein. The anti-SIAH2 therapy is highly effective in inducing oxidative stress, altered mitochondrial bioenergetics, and apoptosis in a subset of invasive and metastatic cancer lines expressing insufficient levels of Survivin. Our results suggest that, anti-SIAH2 strategy reveals novel oncogenic K-RAS-dependent cancer cell vulnerability in stress response that can be exploited in future for anti-SIAH2 therapy alone, or in combination with other anti-proliferative treatment to effectively combat KRAS-dependent cancers. Citation Format: Monicah M. Njogu, Caroline Lee, Jamie Eisner, Minglei Bian, Robert Vansciver, Amy H. Tang. Increased cellular stress and apoptosis induced by anti-SIAH2 therapy in human cancer cells with oncogenic K-RAS addiction. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3498.

Abstract 2487: Nicotine-mediated regulation of Sox2 and stemness in non-small cell lung cancer

Abstract Lung cancer is the leading cause of cancer deaths in both men and women, and is highly correlated with cigarette smoking. Nicotine, the addictive component of tobacco smoke, cannot initiate tumors, but can promote proliferation, migration, and invasion of cells in vitro and promote growth and metastasis of tumors in vivo. More recently, nicotine has been implicated in the promoting self-renewal of stem-like side-population cells from lung cancers. This subpopulation of cancer stem-like cells has been implicated in tumor initiation, generation of entire heterogeneous tumor population, metastasis, dormancy, and drug resistance. We have shown that nicotine promotes the expression of genes that promote stemness, such as Stem Cell Factor (SCF/c-kit ligand), in a specific receptor dependent fashion. Here we demonstrate that nicotine can induce the expression of embryonic stem cell factor Sox2, which is indispensable for self-renewal and maintenance of stem cell properties in non-small cell lung adenocarcinoma cells. High level of Sox2 expression also correlates with poor patient survival outcome in lung cancer, and metastatic lung adenocarcinoma shows higher levels of Sox2 expression than primary tumor or normal tissue, implicating Sox2 in this disease. Nicotine was found to induce Sox2 in A549 cells after 18 and 24 hours of stimulation, an effect which was diminished by 48 hours. This was seen at the mRNA and protein level as assessed by transient transfection and luciferase assay, real-time PCR, western blot, and immunofluorescence respectively. Experiments were conducted to elucidate the mechanism by which nicotine induces Sox2; it was found that siRNA mediated depletion of α7 nicotinic acetylcholine receptor (nAChR), the Hippo pathway effector Yap1, or the transcriptional repressor ID1 could abrogate nicotine-mediated induction of Sox2. Further, treatment with inhibitors of Src kinase or PI3K additionally abrogated the effect, suggesting these proteins play a role in nicotine-mediate induction of Sox2. Interestingly, nicotine could induce Sox2 expression in human mesenchymal stem cells (hMSCs) after 24 hours of stimulation as seen by immunofluorescence and real-time PCR, and this effect was abrogated by treatment with inhibitors to Src kinase, PI3K, Yap1, or α7 nAChR. Additionally, studies are under way to elucidate whether e-cigarettes or flavored tobacco, newer and less studied alternatives to traditional cigarettes, have similar effects on stemness. These studies can be expected to have a direct impact on our understanding of the molecular mechanisms involved in the initiation, growth, and metastasis of non-small cell lung cancer, especially in smokers and tobacco users. Citation Format: Courtney Schaal, Srikumar Chellappan. Nicotine-mediated regulation of Sox2 and stemness in non-small cell lung cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 2487.

Nicotine-Mediated Regulation of Nicotinic Acetylcholine Receptors in Non-Small Cell Lung Adenocarcinoma by E2F1 and STAT1 Transcription Factors

Cigarette smoking is the major risk factor for non-small cell lung cancer (NSCLC), which accounts for 80% of all lung cancers. Nicotine, the addictive component of tobacco smoke, can induce proliferation, migration, invasion, epithelial-mesenchymal transition (EMT), angiogenesis, and survival in NSCLC cell lines, as well as growth and metastasis of NSCLC in mice. This nicotine-mediated tumor progression is facilitated through activation of nicotinic acetylcholine receptors (nAChRs), specifically the α7 subunit; however, how the α7 nAChR gene is regulated in lung adenocarcinoma is not fully clear. Here we demonstrate that the α7 nAChR gene promoter is differentially regulated by E2F and STAT transcription factors through a competitive interplay; E2F1 induces the promoter, while STAT transcription factors repress it by binding to an overlapping site at a region -294 through -463bp upstream of the transcription start site. Treatment of cells with nicotine induced the mRNA and protein levels of α7 nAChR; this could be abrogated by treatment with inhibitors targeting Src, PI3K, MEK, α7 nAChR, CDK4/6 or a disruptor of the Rb-Raf-1 interaction. Further, nicotine–mediated induction of α7 nAChR was reduced when E2F1 was depleted and in contrast elevated when STAT1 was depleted by siRNAs. Interestingly, extracts from e-cigarettes, which have recently emerged as healthier alternatives to traditional cigarette smoking, can also induce α7 nAChR expression in a manner similar to nicotine. These results suggest an autoregulatory feed-forward loop that induces the levels of α7 nAChR upon exposure to nicotine, which enhances the strength of the signal. It can be imagined that such an induction of α7 nAChR contributes to the tumor-promoting functions of nicotine.

Utility of liquid biopsies to assess circulating tumor DNA in patients with lung adenocarcinoma.

e23097Background: Liquid biopsies were used to assess genomic alterations in plasma circulating tumor DNA (ctDNA) from patients with non-small cell lung adenocarcinoma (NSCLC), and therapeutic outc...

Metástasis coroidea como signo de presentación de un adenocarcinoma de pulmón con respuesta a afatinib

Case reportWe present the case of a 55-year-old man with a non-small cell lung adenocarcinoma, who presented with choroidal metastasis. The patient showed a decrease in visual acuity. His evaluation revealed unilateral choroidal metastasis secondary to carcinoma of the lung. The patient received afatinib with complete regression of choroidal metastasis after one year follow-up. DiscussionChoroidal metastasis may be the initial sign of lung cancer. This case highlights the importance of a thorough systemic evaluation in patients with choroidal tumours. Afatinib was effective against choroidal metastasis of a lung adenocarcinoma with EFGR mutation.

Anticancer screening of medicinal plants growing in the Northern region of Saudi Arabia

Background: Desert condition in which plants grow and survive and altering biochemical properties of plants are known to increase the secondary metabolite induction in a variety of medicinal plants. Cancer is a terrible disease and fighting it is of abundant significance to public health. Aims and Objective: To search for new compounds with cytotoxic activity isolated from natural source. In this article, we studied the anticancer properties of three plants (Farsetia aegyptia, Lactuca serriola, and Santolina chamaecyparissus) growing under desert condition of the Northern Region of Saudi Arabia. Materials and Methods: Hydromethanol extracts of the plants were investigated in vitro against four different cell lines. Anticancer activities were assayed with standard 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) colorimetric procedure against A549, HCT116, Hep G2, and MCF7 cell lines. Result: From the analysis, it was found that Lactuca and Santolina sp. showed 71% and 66% cytotoxic activity against Hep G2 and inhibited cell line at 100 µg/mL tested dose; F. aegyptia showed 74% cytotoxic activity against A549 and inhibited cell line at 100 µg/mL. But, the three plants did not show much anticancer activity against HCT116 cancer cell line. Methanol extracts were subjected to gas chromatography-mass spectrophotometry (GC-MS) in order to evaluate the chemical constituents of these plants. Conclusion: It can be concluded that, in F. aegyptia, L. serriola, and S. chamaecyparissus, which grow undrer dry desert condition of Northern Borders Region, the stress condition makes the plant accumulate active compounds that possess anticancer properties against nonsmall cell lung adenocarcinoma and human hepatocellular liver carcinoma.

MicroRNA-7 Compromises p53 Protein-dependent Apoptosis by Controlling the Expression of the Chromatin Remodeling Factor SMARCD1

We previously demonstrated that the epidermal growth factor receptor (EGFR) up-regulated miR-7 to promote tumor growth during lung cancer oncogenesis. Several lines of evidence have suggested that alterations in chromatin remodeling components contribute to cancer initiation and progression. In this study, we identified SMARCD1 (SWI/SNF-related, matrix-associated, actin-dependent regulator of chromatin, subfamily d, member 1) as a novel target gene of miR-7. miR-7 expression reduced SMARCD1 protein expression in lung cancer cell lines. We used luciferase reporters carrying wild type or mutated 3'UTR of SMARCD1 and found that miR-7 blocked SMARCD1 expression by binding to two seed regions in the 3'UTR of SMARCD1 and down-regulated SMARCD1 mRNA expression. Additionally, upon chemotherapy drug treatment, miR-7 down-regulated p53-dependent apoptosis-related gene BAX (BCL2-associated X protein) and p21 expression by interfering with the interaction between SMARCD1 and p53, thereby reducing caspase3 cleavage and the downstream apoptosis cascades. We found that although SMARCD1 sensitized lung cancer cells to chemotherapy drug-induced apoptosis, miR-7 enhanced the drug resistance potential of lung cancer cells against chemotherapy drugs. SMARCD1 was down-regulated in patients with non-small cell lung cancer and lung adenocarcinoma cell lines, and SMARCD1 and miR-7 expression levels were negatively correlated in clinical samples. Our investigation into the involvement of the EGFR-regulated microRNA pathway in the SWI/SNF chromatin remodeling complex suggests that EGFR-mediated miR-7 suppresses the coupling of the chromatin remodeling factor SMARCD1 with p53, resulting in increased chemo-resistance of lung cancer cells.

Classification of Non-Small Cell Lung Cancer Using Significance Analysis of Microarray-Gene Set Reduction Algorithm.

Among non-small cell lung cancer (NSCLC), adenocarcinoma (AC), and squamous cell carcinoma (SCC) are two major histology subtypes, accounting for roughly 40% and 30% of all lung cancer cases, respectively. Since AC and SCC differ in their cell of origin, location within the lung, and growth pattern, they are considered as distinct diseases. Gene expression signatures have been demonstrated to be an effective tool for distinguishing AC and SCC. Gene set analysis is regarded as irrelevant to the identification of gene expression signatures. Nevertheless, we found that one specific gene set analysis method, significance analysis of microarray-gene set reduction (SAMGSR), can be adopted directly to select relevant features and to construct gene expression signatures. In this study, we applied SAMGSR to a NSCLC gene expression dataset. When compared with several novel feature selection algorithms, for example, LASSO, SAMGSR has equivalent or better performance in terms of predictive ability and model parsimony. Therefore, SAMGSR is a feature selection algorithm, indeed. Additionally, we applied SAMGSR to AC and SCC subtypes separately to discriminate their respective stages, that is, stage II versus stage I. Few overlaps between these two resulting gene signatures illustrate that AC and SCC are technically distinct diseases. Therefore, stratified analyses on subtypes are recommended when diagnostic or prognostic signatures of these two NSCLC subtypes are constructed.

Abstract B46: The use of quantitative proteomics to increase the knowledge about metabolic symbiosis in a tumor microenvironment

Abstract For more than a century, it is known that in a solid tumor, malignant cancer cells co-reside with stromal cells including fibroblasts and immune cells. In addition, changes in metabolic requirements of malignant cells are necessary to cover their high proliferative rate. Although it has been demonstrated that complementary metabolic functions of tumor cells and their supporting local stroma (e.g. cancer-associated fibroblasts, adipocytes) contribute to the progression of cancer, less is known about the metabolic needs of immune cells in this local tumor microenvironment. However, in the context of cancer, immune cells, including tumor-associated macrophages (TAMs) are of high interest since it is known that these cells are able to influence tumor progression by allowing the establishment of an attractive environment for tumor growth and metastasis. Studying the expression profiles of proteins involved in metabolic processes of these cells might be a good strategy to elucidate this question. In this study, we applied quantitative proteomics to study the differences in protein expression of TAMs vs. control macrophages (both bone marrow-derived and peritoneal macrophages) isolated from an in vivo murine model for non-small cell lung adenocarcinoma. In our peptide-centric two dimensional liquid chromatography (LC)- tandem mass spectrometry (MS/MS) setup where Tandem Mass Tag (TMT) labeling is applied, we were able to identify 1250 unique proteins, of which 1178 proteins were quantified with all six TMT reporter ions. Interestingly, analysis of the differentially expressed proteins indicate that several Gene Ontology terms regarding metabolic pathways including glutathione metabolic processes (p= 3.47 e-14) and pentose phosphate shunt (p= 2.29 e-8) were enriched. These data suggest that TAMs can reprogram their metabolism in order to survive to the oxidative stress existing in this kind of microenvironment. In conclusion, our results illustrate that proteome analysis is a good strategy to obtain insights in the biology of tumor-associated macrophages. Citation Format: Evelyne Maes, Mathias Wenes, Dirk Valkenborg, Inge Mertens, Hans Prenen, Massimiliano Mazzone, Geert Baggerman. The use of quantitative proteomics to increase the knowledge about metabolic symbiosis in a tumor microenvironment. [abstract]. In: Proceedings of the AACR Special Conference: Metabolism and Cancer; Jun 7-10, 2015; Bellevue, WA. Philadelphia (PA): AACR; Mol Cancer Res 2016;14(1_Suppl):Abstract nr B46.

Abstract C170: Preclinical evaluation of mirvetuximab soravtansine (IMGN853) combination therapy in ovarian cancer xenograft models

Abstract Background: IMGN853 is a folate receptor α (FRα)-binding antibody-drug conjugate (ADC) that utilizes the potent tubulin-targeting maytansinoid, DM4, as its cytotoxic agent. FRα is highly expressed in many solid tumors, particularly epithelial ovarian cancer (EOC), endometrial cancer and non-small cell lung adenocarcinoma. IMGN853 is currently being evaluated as monotherapy in FRα-positive solid tumors in a Phase 1 trial (NCT01609556), with encouraging results recently reported in 17 evaluable patients treated at 6.0 mg/kg adjusted ideal body weight (AIBW) with platinum-resistant EOC (Moore K et al, 2015). Methods: EOC cell line xenograft models plus EOC patient derived xenograft (PDX) models that had FRα expression representative of patients enrolled in the Phase 1 trial were used to assess IMGN853 single agent and combination therapy activity. Anti-cancer therapies used in EOC were assessed. Results from studies with bevacizumab (Bev), carboplatin and pegylated liposomal doxorubicin (PLD) are reported herein. Results: IMGN853 plus Bev was assessed in multiple models including OV90 and IGROV-1 EOC cell line xenografts and a platinum-resistant EOC PDX model, and was consistently more active than either agent alone. In most studies, monotherapy IMGN853 or Bev was active with few partial or complete regressions. In contrast, combination IMGN853 + Bev was highly active, with a majority of the animals having partial or complete tumor regression. The combination activity was substantially more than additive and studies to understand the mechanism(s) responsible for the enhanced activity are ongoing. Combination carboplatin + IMGN853 was more active than carboplatin + paclitaxel in OV90 EOC xenografts. The addition of Bev to carboplatin + paclitaxel enhanced activity compared to carboplatin + paclitaxel. Carboplatin + IMGN853 was more efficacious than the triple combination of carboplatin + paclitaxel + Bev. Carboplatin + IMGN853 + Bev was the most active combination with all mice having tumors that completely regressed. Finally, the combination of PLD and IMGN853 was highly active in a platinum-resistant EOC PDX model, and much more active than PLD or IMGN853 alone. All combinations with IMGN853 described above were well tolerated. Conclusion: Combination therapy efficacy of IMGN853 with Bev was substantially more than additive in multiple models of platinum resistant EOC. Combination IMGN853 + PLD is more efficacious than either monotherapy and combination IMGN853 + carboplatin is more efficacious than carboplatin + paclitaxel in the models studied. Addition of Bev to the carboplatin + IMGN853 combination further enhanced activity. Studies to understand the mechanism(s) responsible for the enhanced combination activity are under way. The efficacy observed in these models suggests that IMGN853 in combination with PLD, or Bev and/or carboplatin may be promising regimens to evaluate in clinical trials of EOC both in the relapsed and upfront settings. A phase1b clinical study assessing doublet combinations of IMGN853 with PLD, Bev and carboplatin in relapsed EOC is planned for 2015. Citation Format: Jose F. Ponte, Jennifer Coccia, Leanne Lanieri, Rabih Gabriel, Jan Pinkas, Rodrigo Ruiz-Soto. Preclinical evaluation of mirvetuximab soravtansine (IMGN853) combination therapy in ovarian cancer xenograft models. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2015 Nov 5-9; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2015;14(12 Suppl 2):Abstract nr C170.

Abstract B170: Discovery of tubulysin payloads for antibody drug conjugates with potent in vitro activity and in vivo efficacy in solid tumor models

Abstract Antibody drug conjugates (ADCs) combine the specificity of antibodies with the potency of small molecule cytotoxic drugs and have the potential to provide significant efficacy as a treatment for cancer. The objective of this work was to identify potent new cytotoxic ADC payloads that can be used to target diverse tumor types. Here we report for the first time the discovery of fully synthetic tubulysin payloads which belong to a class of highly cytotoxic natural products that disrupt the cellular microtubule network leading to apoptosis of tumor cells. Our fully synthetic tubulysin payloads are comprised of: (i) a tubulysin warhead that displays pM potency, (ii) a protease cleavable amino-acid sequence and (iii) a tether bearing a reactive maleimide group. Tubulysin-based ADCs were generated via site-specific conjugation of these payloads to cysteines engineered into antibodies against cancer antigen target oncofetal protein 5T4. The resulting ADCs showed potent in vitro cell killing and in vivo efficacy in multiple solid tumor xenograft models including prostate cancer, non-small cell lung adenocarcinoma, breast cancer and gastric carcinoma. Furthermore, specific structural features of the tubulysin warhead, linker design and antibody engineering were shown to impact the overall in vitro and in vivo properties of the ADCs. Thus, these synthetic tubulysin payloads represent novel microtubule network disrupting compounds that display potent preclinical anti-tumor activity as an ADC that could be advanced to the clinic. Citation Format: Dorin Toader, Jay Harper, Chris Lloyd, Rose Marwood, David Bannister, Shenlan Mao, Cui (Tracy) Chen, Haihon (Helen) Zhong, Vahe Bedian, Fengjiang Wang, Lakshmaiah Gingipalli, Melisa Vasbinder, Pamela Thompson, Ryan Fleming, Byniam Bezabeh, Nazzareno Dimasi, Changshou Gao, Adeela Kamal. Discovery of tubulysin payloads for antibody drug conjugates with potent in vitro activity and in vivo efficacy in solid tumor models. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2015 Nov 5-9; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2015;14(12 Suppl 2):Abstract nr B170.