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Developmental Projects

Where You Are:

The developmental component of the SPORE supports two funding mechanisms for lung cancer research: Developmental Research Projects and Career Development Projects.  These awards are available to Moffitt affiliated researchers as described below under the individual awards.

Developmental Research Project Awards provides financial support for faculty members (Associate Professor level or higher) interested in developing (or further developing) research programs in lung cancer.  There are two $75,000 awards distributed for a one year period.  The primary criteria for these awards are the quality of the research proposal and its relevance to lung cancer and to this SPORE.    

Career Development Awards provides financial support for early career faculty members (Assistant Professor level or equivalent) interested in developing (or further developing) research programs in lung cancer.  There are two $50,000 awards distributed for a one year period.  The primary criteria for these awards is the applicant’s potential for a career as an independent investigator in translational research of lung cancer.

A “Call for Applications” is announced on an annual basis.  For more information on the application process or deadlines, please contact Carol Ulge at 813-745-3886 or carol.ulge@moffitt.org.

Questions about this award mechanism should be addressed to Dr. Doug Cress at douglas.cress@moffitt.org.  

Information on projects currently or previously funded is included below.

Career Developmental Projects

Career Development Awards provide financial support for early career faculty members (Assistant Professor level or equivalent) interested in developing (or further developing) research programs in lung cancer.  There are two $50,000 awards distributed for a one year period.  The primary criteria for these awards is the applicant’s potential for a career as an independent investigator in translational research of lung cancer.

1. Sept 2010 - Aug 2012

Sept 2011 - Aug 2012
Career Developmental Projects (2011-2012 - Year 4)
The SPORE funded the following career developmental projects:

  • Developing molecularly targeted combination therapies in a preclinical mouse model of lung cancer - Dr. Vrushank Dave
  • Molecular imaging probes for lung cancer intraoperative guidance - Dr. David Morse
  • Probing Aberrant Signaling Circuitries in Non-Small Cell Lung Cancer with Kinase Inhibitors Using A Combination of Chemical and Phosphoproteomics - Dr. Uwe Rix

Developing molecularly targeted combination therapies in a preclinical mouse model of lung cancer - Dr. Vrushank Dave

Abstract

The main objective of this research proposal is to develop therapies for patients with lung cancer associated with aberrant PI3K/mTOR and KRAS signaling that become chemo/radio-resistance and predict poor prognosis with conventional therapies. To this end, we will develop pre-clinical mouse models of lung cancer associated with hyper-activated AKT/mTOR and oncogenic KRAS pathways, and ascertain the mechanisms of action of combination therapies with PI3K/mTOR and pan-RAF inhibitors. PI3K/mTOR pathway is an ideal therapeutic target as mTOR is a hub for growth stimuli that promote tumor initiation, maintenance, apoptotic resistance, epithelial-mesenchymal transition (EMT) and metastasis. The PTEN tumor suppressor is the primary controller of the PI3K/mTOR pathway. The central hypothesis is that mTOR hyper-activation due to PTEN loss in the presence of oncogenic KRAS causes aggressive lung tumors associated with EMT, and that, our inhibitors of mTOR- and KRAS-pathway in combination, will reduce tumor growth, cause apoptosis and inhibit EMT. We will generate lung epithelial specific inducible conditional deletion of PTEN gene and activate mutant KRAS-G12D and PI3K/mTOR pathway utilizing CCSP and SPC-promoters in the conducting and peripheral airways respectively, so as to cause lung cancers of distinct cell origins. We will: 1) Establish a mouse models of lung cancer associated with hyper-activated mTOR and KRAS and assess the efficacy of PI3K/mTOR and KRAS pathway-inhibitors for anti-growth and pro-apoptotic effects on tumors in vivo. 2) Elucidate how PI3K/mTOR and KRAS pathway-inhibitors influence transcription activation of master regulators of EMT that play critical roles in migratory and invasive properties of cancer cells, increasing the metastatic potential. Upon completion of this proposal we expect to define the role of selective inhibitors of PI3K/mTOR and KRAS pathway in treating lung cancers and the mechanism by which they modulate EMT. The proposed models provide an easy entrée to develop novel EMT-targeted therapy, and more importantly, test and validate safer and effective targeted combination therapies for clinical trials in lung cancer patients with KRAS lesions associated with hyperactivated PI3K/mTOR pathway that are ongoing as part of the Moffitt Lung SPORE clinical trials.

Molecular imaging probes for lung cancer intraoperative guidance - Dr. David Morse

Abstract
A major goal in molecular imaging of cancer is to deliver imaging contrast to tumor with high selectivity. Targeting the cell surface is appealing because such agents can be more structurally diverse, compared to agents that must cross the plasma membrane. In cancers, antibody-based surface targeting agents have been used to successfully deliver targeted therapy. However, compared to small peptide ligands, antibodies have less favorable pharmacokinetic (PK) profiles, such as increased circulation and clearance times, decreased rates of extravasation, decreased penetration into tumors and increased non-specific interactions. The current work seeks to develop a peptide imaging probe against a bona fide lung cancer cell-surface marker for the purpose of margin detection during surgery. Such probes can enable the real-time detection and thus improve removal of residual disease in the surgical cavity, which we expect will reduce recurrence of disease and increase patient survival. A second goal of this study is to identify and validate novel lung cancer markers that cover all lung cancer types, allowing the future development of additional probes.
The delta-opioid receptor (dOR) is known to be overexpressed in some lung cancers and is not expressed in normal lung. We have conjugated a fluorochrome to a synthetic peptide dOR antagonist (Dmt-Tic). The resulting probe retained high in vitro binding affinity and selectivity for the dOR, and high in vivo tumor selectivity with favorable PK and biodistribution profiles. The current proposal will include two independent aims. Aim 1 will develop a fluorescent probe by conjugating a longer wavelength fluorochrome (IR800CW, LICOR) to Dmt-Tic for increased detectability in tissue. The second aim will use gene expression profiling and immunohisto-chemistry to identify and validate additional cell-surface targets to enable coverage of 100% of lung cancers.
This study will be leveraged by biomarker discovery and imaging approaches we have already developed in other cancers (e.g. pancreatic, melanoma, breast) with the Lung SPORE biomarker discovery efforts, e.g. the construction of a database containing of patient lung tumor sample microarray data, the use of biomarker discovery analyses that have already been developed for lung cancer, and the construction of tissue microarrays containing patient lung cancer tissue samples for validation of marker protein expression.
By the end of this study, we will have developed and pre-clinically characterized a lung cancer targeted fluorescent probe that can be further translated into clinical trials. Additional markers will also be identified and validated that cover 100% of lung cancers and metastases, enabling future development of additional imaging probes. Such advances will allow for intraoperative margin detection of lung cancer, with potential to improve patient survival by increasing the percentage of complete surgical resection without residual disease. In addition to translation of these as intraoperative imaging agents, these probes may also be developed as a platform for delivery of targeted therapies in future studies. Future studies may also use existing patient data retrospectively to characterize these markers for use in diagnosis and prognosis.

Sept 2010 - Aug 2011
Career Developmental Projects (2010-2011 - Year 3)
The SPORE funded two career developmental projects as follows:

  • Phosphorylation Profiling and Shotgun Proteomics to Discover Prognostic Biomarkers for Early Stage Lung Cancer: Dr. John Koomen
  • Role of histone deacetylase 6 and p53 in cisplatin-resistance of lung cancer: Dr. Mary Zhang
Phosphorylation Profiling and Shotgun Proteomics to Discover Prognostic Biomarkers for Early Stage Lung Cancer: Dr. John Koomen

Abstract
The focus of this grant proposal is the detection of prognostic biomarkers for predicting survival outcomes for early stage lung cancer using a combination of phosphorylation profiling and shotgun proteomics. This pilot project would compare data from 10 tumors from early stage non small cell lung cancer patients with good outcomes, defined as curative treatment with no history of relapse or recurrence, with data from 10 tumors from patients with poor outcomes, defined as survival shorter than 18 months. Because the spectrum of lung cancer outcomes includes these points at each end of the scale, the dichotomous comparison of these two groups should yield potential biomarkers for patient assessment.
Candidate biomarkers with be detected and identified using bottom-up discovery based proteomics methods centered around liquid chromatography coupled to tandem mass spectrometry peptide sequencing (LC-MS/MS). The major outcome of an LC-MS/MS experiment is a catalog of peptide identifications obtained by database searching, which can be used to identify their protein of origin as well as specific sites of posttranslational modification. Here, we propose to fractionate tryptic peptides from tissue homogenate with strong cation exchange (SCX) prior to immobilized metal affinity chromatography enrichment of phosphorylated peptides (IMAC). Both the selected phosphopeptides and the remaining unmodified sequences are then analyzed with LC-MS/MS to create a phosphorylation profile and proteome catalog for each tumor specimen. Quantitative analysis of each phosphorylation site will rely on extracted ion chromatograms (EIC), which integrate the total peak area for each peptide ion signal in the sample. Comparison of the averages and standard deviations with statistical verification using p values obtained by t-test will determine signals that differ between patients that have good and poor survival outcomes. Quantitative analysis of the proteome catalog can be achieved with two data analysis methods: EIC and enhanced spectral counting (ESC). The latter method uses the total number of peptides detected from each protein multiplied by the total amount of fragment ion signal in the tandem mass spectrum to enable comparison between sample groups. These complementary quantification tools make full use of the LC-MS/MS data for comparison.

These analyses will produce data to show feasibility and proof of principle for a mass spectrometry based prognostic biomarker discovery platform applied to lung cancer that will support future grant applications. More importantly, the data will be mapped across different signaling pathways for the phosphorylation profiles and across known biological processes for the proteome catalog. These efforts may elucidate common phenotypes for each group of cancer patients which can be used to predict survival outcomes and suggest possible therapeutic targets for the patients with poor survival outcomes. Finally, a set of candidate prognostic biomarkers, including specific proteins and their phosphorylations, will be developed that can be tested in biopsy specimens (either cores or final needle aspirates) using liquid chromatography coupled to multiple reaction monitoring mass spectrometry (LC-MRM) or antibody based methods (e.g. enzyme-linked immunosorbent assays) to determine their predictive value over the spectrum of lung cancer patients’ survival outcomes. This set of experiments can help to define a molecular basis for evaluation of lung cancer patients, which is the first step in the development of personalized treatment regimens determined by aspects of the tumor biology observed in each patient.

Role of histone deacetylase 6 and p53 in cisplatin-resistance of lung cancer: Dr. Mary Zhang

Abstract

With 1.2 million new cases diagnosed every year, lung cancer is the leading cause of cancerrelated mortality in both men and women. Two first line regimens of lung cancer treatment are platinum with taxane and platinum with gemcitabine. One of the major obstacles of these treatments is that patients often develop resistance to platinum. Histone deacetylases (HDACs) are enzymes which remove acetyl moiety from histone and non-histone proteins, thereby being involved in a wide variety of biological processes including transcription regulation, cell growth/differentiation, apoptosis, etc. HDAC inhibitors now hold great promises in cancer treatment that they could restore the chemo-sensitivity in tumor cells, including lung cancer cells. However, the role of these enzymes in chemo-resistance is largely unknown. Our preliminary data show that one of the HDACs, termed HDAC6, acts as a ubiquitin E3 ligase to promote p53 polyubiquitination which leads to p53 degradation that may account for cisplatin resistance. Moreover, we have shown that HDAC6 protein levels positively correlate with cisplatin resistance in a panel of lung cancer cells. Therefore, in this renewed grant, we will further characterize HDAC6 E3 ligase activity by defining the minimal E3 ligase domain, E2 binding site and E3 ligase catalytic center. We will also extend our studies from lung cancer cell lines to lung cancer tissue specimens to examine HDAC6 and p53 expression in platinum-sensitive and platinum-resistant patient samples. The results of our research will identify HDAC6 as a novel therapeutic target in lung cancer and suggest the application of clinically relevant HDAC6-selective inhibitors as adjuvant drugs.

2. Sept 2008 - Aug 2010

Sept 2009 - Aug 2010
Career Developmental Projects (09/10-Year 2)
The SPORE funded two career developmental projects as follows:

  • Role of Activated Ack1/AKT Signaling in Lung Cancer: Nupam Mahajan
  • Role of Histone Deacetylase 6 and p53 in cisplatin-resistance of lung cancer: Mary Zhang

Role of Activated Ack1/AKT Signaling in Lung Cancer: Nupam Mahajan

The AKT/PKB kinase is a key signaling component of one of the most frequently activated pathways in cancer and is a major target of cancer drug development. Most studies have focused on its activation by Receptor Tyrosine Kinase (RTK) mediated Phosphatidylinositol-3-OH kinase (PI3K) activation or loss of Phosphatase and Tensin homolog (PTEN). We have uncovered that growth factors binding to RTKs lead to activation of a non-receptor tyrosine kinase, Ack1 (also known as TNK2), which directly phosphorylates AKT at an evolutionarily conserved tyrosine 176 in the kinase domain. Tyr176-phosphorylated AKT localizes to the plasma membrane and promotes Thr308/Ser473-phosphorylation leading to AKT activation. Expression levels of Tyr176-phosphorylated-AKT and Tyr284-phosphorylated-Ack1 were positively correlated with the severity of disease progression, and inversely correlated with the survival of cancer patients. Thus, RTK/Ack1/AKT pathway provides a novel target for drug discovery.

Role of Histone Deacetylase 6 and p53 in cisplatin-resistance of lung cancer: Mary Zhang

With 1.2 million new cases diagnosed every year, lung cancer is the leading cause of cancer-related mortality in both men and women. Two first line regimens of lung cancer treatment are platinum with taxane and platinum with gemcitabine. One of the major obstacles of these treatments is that patients often develop resistance to platinum. Histone deacetylases (HDACs) are enzymes which remove acetyl moiety from histone and non-histone proteins, thereby being involved in a wide variety of biological processes including transcription regulation, cell growth/differentiation, apoptosis, etc. HDAC inhibitors now hold great promises in cancer treatment that they could restore the chemo-sensitivity in tumor cells, including lung cancer cells. However, the role of these enzymes in chemo-resistance is largely unknown. Here, we have shown that HDAC6 interacts with and deacetylates the DNA mismatch repair proteins, MSH2 and MSH6, which play critical roles in recognition of cisplatin-induced DNA adducts and trigger programmed cell death. Cisplatin is one of the most widely used platinum compounds in lung cancer treatment and we will use it as a representative drug to study the platinum-resistance. We have further shown that knockdown of HDAC6 sensitizes cisplatin-induced cell death. Moreover, HDAC6 level is positively correlated with cisplatin’s IC50 in a panel of lung cancer cell lines. In addition, our data have shown that HDAC6 regulates 6-thioganuine resistance via MSH2, indicating that HDAC6 conferred cisplatin resistance may also via DNA mismatch repair proteins. Overall, our results identify HDAC6 as a novel therapeutic target in lung cancer and suggest the application of clinically relevant HDAC6-selective inhibitors as adjuvant drugs.

Sept 2008 - Aug 2009

Career Developmental Projects (08/09-Year 1)
The SPORE funded three career developmental projects as follows:

  • Role of Novel Methy-H3K9 Binding Protein MPP8 in Lung Cancer Metastasis: Dr. Jia Fang
  • Role of Activated Ack1 in Lung Cancer: Dr. Nupam Mahajan
  • Molecular Epidemiology of Lung Cancer Survivorship: Protease-Related Gene Polymorphisms: Dr. Matthew Schabath

Role of Novel Methy-H3K9 Binding Protein MPP8 in Lung Cancer Metastasis: Dr. Jia Fang

Histone H3K9 methylation is one of the best studied epigenetic modifications that has been linked to a variety of biological processes such as position-effect variegation, heterochromatin formation and transcriptional regulation. The methyl groups on H3K9 serve as binding sites for regulatory proteins which in turn mediate different functional outcomes. To further understand the functions of H3K9 methylation, we identified MPP8 as a novel methyl-H3K9 binding protein and demonstrated that the binding is mediated through its chromodomain in vitro and in vivo. MPP8 expression is elevated in various human lung cancer cells and correlated with cell invasiveness suggesting MPP8 is involved in tumor metastasis. Microarray analysis revealed that MPP8 regulates a large group of genes that are involved in cell adhesion and epithelial-to-mesenchymal transition and the ChIP assays confirmed that MPP8 targets onto the promoter regions of E-cadherin and regulates the expression of this key regulator of cell behavior and tumor progression. To elucidate the molecular mechanism underlying this, we further demonstrated that MPP8 not only specifically interacts with two major euchromatic H3K9 histone methyltransferases GLP/Eu-HMTase1 and ESET/SETDB1, but also the de novo DNA methyltransferase 3A. Together, our results not only uncovers MPP8 as a novel H3K9-methyl binding protein, but also suggest a model by which MPP8 coordinates histone and DNA methylation for tumor suppressor gene silencing which plays an important role in epithelial-to-mesenchymal transition, tumorigenesis and metastasis.

Role of Activated Ack1 in Lung Cancer: Dr. Nupam Mahajan

The AKT/PKB kinase is a key signaling component of one of the most frequently activated pathways in cancer and is a major target of cancer drug development. Most studies have focused on its activation by Receptor Tyrosine Kinase (RTK) mediated Phosphatidylinositol-3-OH kinase (PI3K) activation or loss of Phosphatase and Tensin homolog (PTEN). We have uncovered that growth factors binding to RTKs lead to activation of a non-receptor tyrosine kinase, Ack1 (also known as TNK2), which directly phosphorylates AKT at an evolutionarily conserved tyrosine 176 in the kinase domain. Tyr176-phosphorylated AKT localizes to the plasma membrane and promotes Thr308/Ser473-phosphorylation leading to AKT activation. Mice expressing activated Ack1 specifically in the prostate exhibit AKT Tyr176-phosphorylation and develop prostatic intraepithelial neoplasia (PINs). Further, expression levels of Tyr176-phosphorylated-AKT and Tyr284-phosphorylated-Ack1 were positively correlated with the severity of disease progression, and inversely correlated with the survival of breast and pancreatic cancer patients, suggesting that RTK/Ack1/AKT pathway provides a novel target for drug discovery. Consistent with this data, a small molecule inhibitor of Ack1, 4-amino-5,6-biaryl-furo[2,3-d]pyrimidine not only inhibited Ack1 activation, but was able to inhibit lung cancer cell proliferation by arresting cells in G0/G1 phase of cell cycle. Collectively, evidence indicates that targeting Ack1 kinase in lung cancer patients could be highly effective therapeutic strategy.

Molecular Epidemiology of Lung Cancer Survivorship: Protease-Related Gene Polymorphisms: Dr. Matthew Schabath

Lung cancer remains a leading cause of cancer death for both men and women in the United States. Presently, the 5-year lung cancer patient survival rate is only a dismal 16%. The grim outlook of lung cancer demonstrates the need to advance individualized medicine to improve lung cancer outcomes and survivorship. At present there are limited data on the role of germline variations (i.e., SNPs) on lung cancer outcomes, but association studies have provided intriguing evidence that the same genetic polymorphisms which are implicated in lung cancer risk also modify patient survival and outcome. This career development project has proposed (1) to assess the role of 1,536 SNPs on lung cancer survival in an existing cohort of 723 lung cancer patients and (2) to build the infrastructure to prospectively recruit lung cancer patients to augment the existing cohort. The proposed SNPs were selected a priori based on two or more of the following criteria: frequency = 5%, from a pathway-specific gene (e.g., DNA repair, inflammation, cell cycle, etc), found to be significant in a previous GWAS or association study, and/or has functional significance. This application plans to test the underlying biological hypothesis that germline genetic variations in candidate genes play an important role in lung cancer survival. Translational and integrative epidemiology research has the potential to improve patient outcomes and survival by identifying molecular/genetic markers to predict relevant and important clinical outcomes such as overall survival, progression free survival, recurrence, toxicity, QOL/PS, and pain/fatigue/depression. This will be one of the first large-scale epidemiology studies that was exclusively designed and developed to assess the genetics of lung cancer patient outcomes and survival.

Developmental Research Projects

Developmental Research Project Awards provides financial support for faculty members (Associate Professor level or higher) interested in developing (or further developing) research programs in lung cancer.  There are two $75,000 awards distributed for a one year period.  The primary criteria for these awards are the quality of the research proposal and its relevance to lung cancer and to this SPORE. 

1. Sept 2010 - Aug 2012

Sept 2011 - Aug 2012
SPORE Developmental Projects
Developmental Research Projects (2011-2012 – Year 4)
The SPORE funded the following developmental research projects:

  • Regulation of immune responses in lung cancer by targeting TRAIL receptor - Dr. Dimitry Gabrilovich and Dr. Scott Antonia
  • Targeting DNA damage response kinases in lung cancer - Dr. Alvaro Monteiro

Regulation of immune responses in lung cancer by targeting TRAIL receptor - Dr. Dimitry Gabrilovich and Dr. Scott Antonia

Abstract
Inadequate function of the host immune system is one of the major factors limiting clinical success of cancer immunotherapy. Myeloid-derived suppressor cells (MDSC) play a major role in inhibiting antigen-specific and nonspecific immune responses in most types of cancer. Their major role in the regulation of immune responses in lung cancer has been demonstrated in animal models and in patients with small and non-small cell lung cancer. Moreover, these cells were implicated in promoting tumor angiogenesis, tumor cell invasion and metastases. MDSC are pathologically activated, immature myeloid cells with potent immune suppressive activity. In recent preliminary experiments, we have discovered that MDSC have a substantially higher expression of TNF-related apoptosis-inducing ligand (TRAIL) receptor than their normal counterparts. Importantly, no differences in the expression of other members of this family (Fas/FasL, RANK/RANKL, TNFR) were found. TRAIL and TRAILR are part of the extrinsic death pathway. TRAIL ligates two types of receptors: death receptors, triggering TRAILR1 (DR4) and TRAILR2 (DR5), which lead to cell apoptosis; and decoy receptors, TRAILR3 (DCR1) and TRAILR4 (DCR2), that possibly inhibit this pathway. A large number of different tumor cells, including non-small cell lung cancer (NSCLC), express TRAILR1 and R2. In contrast, normal myeloid cells, although expressing an abundant amount of TRAIL, have a relatively low amount of TRAILR1 or R2 and are resistant to TRAIL induced cell death.

In the presence of cells expressing high levels of TRAIL (myeloid and lymphoid cells), MDSC quickly undergo apoptosis via the caspase 8/caspase 3 pathways. This effect was much stronger than that observed in the control immature myeloid cells from tumor-free mice. After adoptive transfer to recipient mice, MDSC survived poorly in organ containing a large number of myeloid cells (spleen); but the survival of MDSC was much better in lung, liver, or inside tumor. These observations suggest a possible mechanism of high MDSC turnover in spleens of tumor-bearing mice. It also helps to explain why MDSC can accumulate in some organs and in tumor tissues. We have also demonstrated that cross-linking of TRAILR2, with agonistic antibody, did not affect the viability of control immature myeloid cells; but resulted in significant killing of MDSC. This suggested that targeting TRAILR could be a valuable strategy for a selective elimination of MDSC in tumor-bearing hosts.

In this development research project, we propose to explore our novel concept of regulation of MDSC turnover in tissues. We plan to investigate the mechanisms and functional consequences of TRAILR expression on MDSC. We also plan to evaluate the possibility of therapeutic targeting of TRAILR using agonistic antibody. The goal of this approach is to enhance tumor-specific immune responses in lung cancer by eliminating immune suppressive MDSC. In addition, the elimination of MDSC may interfere with tumor agoigenesis and the ability of tumor to metastasize, which would further contribute to the antitumor effect of the therapy. TRAILR1 and R2 antibody have been developed by several large pharmaceutical companies to target these receptors on tumor cells. Several phase I/II clinical trials were performed in patients with NSCLC with some encouraging results. This opens an opportunity to combine antitumor and immune regulatory effects of TRAILR antibodies in lung cancer. Three specific aims are proposed:

  1. To study the mechanisms and functional consequences of TRAILR2 up-regulation in MDSC from mice bearing lung cancers;
  2. To investigate expression of TRAILR1 and R2 in MDSC from patients with NSCLC;
  3. To determine the effect of TRAILR2 targeting therapy on MDSC in mice with lung cancer
This proposal combines the efforts of Dr. Gabrilovich, who will lead the experimental studies, and Dr. Antonia, who will supervise experiments related to clinical material. The ultimate goal is to develop a full-length SPORE proposal focused on translational study of TRAILR targeting in lung cancer. This future proposal will include abasic biology study of MDSC and the immune responses in tumor-bearing mice, and several clinical trials: one, a phase II trial in patients with NSCLC, aiming to investigate the effect of TRAILR2 on MDSC and non-specific immune responses; and the second, a larger follow-up trial to investigate the combination of TRAILR2 targeting with cancer vaccine.
 

Targeting DNA damage response kinases in lung cancer - Dr. Alvaro Monteiro

Abstract

Eukaryotic cells have evolved an intricate system to resolve damage to DNA and preventing its transmission to daughter cells. This system, collectively known as the DNA damage response (DDR) network includes a large number of proteins responsible for sensing DNA damage, promoting repair, and coordination with cell cycle progression. It has been demonstrated that perturbations in this network can lead to cancer and therefore proteins in this network may constitute excellent candidate targets for therapy.

Recently, we have finalized a map of protein-protein interactions centered on the BRCT domain, a protein modular domain critical for relaying signals in the DDR. This high confidence protein-protein interaction network was generated by combining literature curation, seven parallel yeast two hybrid (Y2H) screens, and tandem affinity purification coupled to mass spectrometry (TAP/MS) of seven protein baits in the presence and absence of ionizing radiation. It contains 718 proteins, of which 19 are protein kinases, all of which target serine/threonine residues. Differently from growth factor receptor signaling pathway kinases that have been intensively studied as targets in therapy, kinases in the DDR have not been systematically explored as potential single agent targets or as sensitizers to therapy.

In this pilot, we propose a systematic screen of protein serine-threonine kinases contained in our empirically determined network in lung cancer cell lines. We contend that inhibition of kinases in this network have a high prior probability of acting as effective sensitizers to currently used chemotherapy drugs. Tyrosine kinase targets have been recently developed based on our emerging knowledge of oncogene addiction in growth factor receptor pathways. On the other hand targeting serine/threonine kinases has not been fully explored yet.

We plan to achieve the following specific aims: To determine the extent to which inhibition of the BRCT network kinases affect lung cancer cells. We hypothesize that these kinases affect the ability of lung cancer cell lines to respond to DNA damage. We will generate lung cancer cell lines depleted (by shRNAs) in each of nineteen kinases identified in the BRCT networks and determine the effect of depletion on their sensitivity to cisplatin and paclitaxel. We expect to identify kinases whose inhibition will lead to sensitization to current chemotherapy drugs. To determine the prevalence of changes in levels in tumors and in cell lines of the most promising kinases identified in the inhibition assays. We hypothesize that kinases showing the most dramatic changes in levels in a series of lung cancers, as measured by western blot, qPCR or automated quantitative analysis of in situ protein expression in tissue microarrays (AQUA) will be the most effective targets of inhibition. Thus we will focus on the most promising leads from the first aim.

This pilot has a main overall objective to develop a framework for a rigorous fast assessment of a protein target as a sensitizer of chemotherapy. Ultimately, targets with promising results can be prioritized to more in-depth pre-clinical studies and clinical trials. The use of systems biology approaches using large datasets of empirically-determined interactions in the DNA Damage Response to identify potential sensitizers of chemotherapy is highly innovative. 

Sept 2009 - Aug 2010
SPORE Developmental Projects
Developmental Research Projects (2010-2011 – Year 3)

The SPORE funded the following developmental research projects:
  • Identification and characterization of stem cells in non-small cell lung carcinoma: Dr. Srikumar Chellappan
  • Validation of CD44/ß1 integrin axis in mediating metastasis and drug resistance in lung cancer: Dr. Lori Hazlehurst
  • Discovery of genetic polymorphisms accounting for reduced lung cancer susceptibility among Puerto Ricans: Dr. Teresita Muñoz-Antonia

Identification and characterization of stem cells in non-small cell lung carcinoma: Dr. Srikumar Chellappan

Abstract

Stem cells from various cancers have been identified based on variety of surface markers, which vary among different types of tumors (1) (2). The stem-like properties of such cells are generally verified by their ability to maintain stemness in defined media, to for three-dimensional colonies in appropriate matrices as well as to give rise to tumors in immunodeficient mice from a very low number of cells. Many of these features distinguish the stem cell concept originally described in myeloid leukemias to the cancer stem cell concept now held for solid tumors (3). The cell surface markers generally associated with breast cancers include high levels of CD44 and low levels or absence of CD24. In addition, may cancer stem cells have been found to have higher levels of CD133. Unfortunately, the expression pattern of these markers is not universal and cannot be used to identify cancer stem cells in other organs (1). In this context, Identification of cancer stem cells in nonsmall cell lung cancer has been a challenge, though such cells were first proposed to be present as early as 1982. Even though CD133 has been proposed to be a viable marker for lung cancer stem cell (4), this does not appear to be universally true. Thus, identification of cancer stem cells associated with non-small cell lung cancer has remained a challenging proposition.

One additional feature of cancer stem cells is their resistance to a variety of drugs (5). To a certain extent, this is due to the overexpression of adenosine triphosphate-binding cassette (ABC) transporters that efflux drugs out of the cell (6) . In the absence of specific markers, the presence of ABC transporters has been used as a surrogate marker for cancer stem cells, including those for the lung (1, 2). The presence of these transporters can be assessed by their ability to efflux the Hoechst33342 dye; such cells can be identified as a side population in FACS analysis (7) (8). Such Side population cells show features of cancer stem cells including self renewal activity, differentiated progeny production, tumorigenicity and expression of stem cell specific genes (9). It has been shown that ABCG2, a member of the ABC transporter superfamily, is an important determinant of the Side population phenotype and is an effective marker for stem like cells. Another marker that has been used in conjunction with ABCG2 is the aldehyde dehydrogenase (ALDH) gene (2). Cancer stem cells identified using side population analysis are subjected to additional in vitro assays to establish their stemness; these include sphere-formation assays, serial colony forming unit assays and label retention assays (10).
Since there are no established markers for lung cancer stem cells, and since CD133 has not been a consistent marker for NSCLC stem cells, we attempted to identify such cells from cultured NSCLC lines by isolating side population and assessing their stemness (11). As shown in the preliminary results, we have succeeded in isolating such cells from H1650 cell line and have conducted preliminary characterization of such cells. We find that such side population cells form spheroids in culture while main population cells don’t, they form colonies in matrigel more efficiently and are more resistant to chemotherapeutic agents. Interestingly, these side population cells could form angiogenic tubules in matrigel and expressed CD31, showing that they are capable of acquiring features of many cell types present in a tumor. At the molecular levels, stemness of these cells required EGFR function as well as activity of ?-arrestin-1 protein. SP cells showed overexpression of c-Kit , SCF as well as transcription factors involved in stemness such as Oct4, Nanog, and Sox2. Based on these observations, we will attempts to characterize the molecular events that mediate stemness of NSCLC side population cells and assess whether targeting EGFR, Sox2 or ?-arrestin-1 will reduce stemness of these cells. The Specific Aims for this study are:
  1. To elucidate the downstream mediators of EGFR that contributes to stemness
  2. To understand the relative contribution of Oct4, Sox2 and Nanog to stemness of NSCLC cells
  3. To elucidate the molecular events that promote the growth of angiogenic tubules from stem-like cells
We believe that these studies will shed light on the molecular mechanisms underlying stemness of NSCLC cells. These studies will enable us to submit a RO1 grant in the very near future.
 

Validation of CD44/ß1 integrin axis in mediating metastasis and drug resistance in lung cancer: Dr. Lori Hazlehurst

Abstract

Lung cancer remains the most deadly cancer and the identification and validation of novel targets are required to improve patient outcome. Recently, it has become clear that tumors evolve in the context of the tumor microenvironment(1). For solid tumors it is becoming apparent that cancer associated fibroblasts (CAF’s) are critical for the progression and metastatic potential of the disease. Furthermore, several recent studies have suggested that it is the bone marrow derived mesenchymal stem cells (MSC’s) located within the stroma that drive metastasis and tumorgenesis(2). MSC’s provide a rich deposit of components of the extracelullar matrices including, fibronectin, collagen and hyaluronic acid (HA). For the past 10 years we have been investigating the role of ß1 integrin in mediating drug resistance in hematopoietic malignancies (3-10). More recently, we showed that HYD1, an integrin antagonist, induces cell death and reverses resistance to standard therapy associated with a co-culture bone marrow stroma model in multiple myeloma (11). Moreover recent data generated in our laboratory indicates that HYD1 binds directly to recombinant CD44v but requires in intact CD44/a4ß1 for inducing cell death in myeloma cell lines (manuscript in preparation). In addition to our compound, currently multiple agents that target ß1 integrin or associated signaling pathways are available that could be rapidly transitioned into the clinic. These agents include Tysarbri a humanized a4 integrin antibody. Tysarbri has been approved for multiple sclerosis, and is currently in phase I trials in multiple myeloma(12). As shown in Figure 1, downstream of ß1 integrin signaling include available agents, which block FAK, Src and integrin linked kinase (ILK)(13, 14). In this proposal we will initially validate the target (CD44 and ß1 integrins) using in vivo metastatic models and primary lung cancer patient specimens. To this end, the goal of specific aim 1 of this grant is to verify whether CD44 and/or ß1 integrin is required for engraftment of lung cancer cell lines into the bone using an intracardiac injection in vivo model of lung metastasis. The goal of specific aim 2, is to determine on a small set of patient derived specimens whether expression of ß1 integrin and/or CD44 correlate with relapse in metastatic sites and or time to death following surgical resection of the primary tumor. Finally, in specific aim 3, of this proposal we will test our novel inhibitor which is a cyclized peptidomimetic of HYD1. The cyclized peptidomimetic of HYD1 is 30 fold more potent compared to the linear counterpart. If our data in specific aim 1 or 2, support that ß1 and/or CD44 protein expression are a determinant of metastatic lung cancer, we will work with drug companies to gain access to compounds that will be readily available to translate to the clinic such as the Pfizer FAK inhibitor. However, we argue that the innovation and novelty of our project is dependent on the target and the fact that we have developed a novel inhibitor. Our inhibitor is distinct from RGD containing peptides. For these reasons we think it is necessary to focus and continue to provide pre-clinical data on the efficacy of this novel inhibitor for securing Florida State Grants as well as NCI based funding. Thus in specific aim 3 of this proposal we will test the potency of our newly synthesized cyclic peptidomimetic c-HYD1 as a single agent and in combination with standard therapy used to treat lung cancer.

Discovery of genetic polymorphisms accounting for reduced lung cancer susceptibility among Puerto Ricans: Dr. Teresita Muñoz-Antonia

Abstract

Lung cancer is the leading cause of cancer death among Hispanic men, and the third most commonly diagnosed cancer among Hispanic men and women. Hispanic/Latinos in the USA are very diverse group in terms of nationality, culture and socio-economic status that share a common language. Few studies have addressed cancer incidence and mortality in the different Hispanic subgroups. For many cancer types, Hispanics living in their country of origin have a lower incidence of cancer than Non-Hispanic Whites living in the USA. Two recent studies comparing the lung cancer incidence and mortality of Puerto Ricans living in PR to those living in the USA found that that lung cancer incidence is lower in Puerto Ricans living in the island compared to Puerto Ricans living in mainland USA, suggesting that lifestyle and environmental factors might play a role in this difference. Interestingly, Puerto Ricans living in mainland USA have a lower incidence than non-Hispanics Whites living in mainland USA, suggesting that in addition to lifestyle and environmental factors, there might be a genetic component for the lower incidence of lung cancer in Puerto Ricans. It is not clear what is contributing to these differences, but it is likely that multiple factors are involved, stressing the need to study the effect of genetic polymorphisms in disease risk in the context of environmental exposure.

Since lung cancer is a complex disease influenced by multiple genes and environmental factors, especially smoking, understanding the interplay of gene and environmental factors could provide novel and practical opportunities for screening, prevention, and early detection. Although several studies have published data assessing the association between single nucleotide polymorphisms (SNPs) and lung cancer risk in Hispanics these studies do not distinguish among different Hispanic groups, nor do they take into consideration that genetically Hispanics are descendents of indigenous Americans, European and African populations. At present, however, little is known on how genetic variability and environmental risk factors may differentially influence lung cancer risk between Hispanics in the US versus Hispanics in the PR. In this pilot project we propose to (1) establish an infrastructure of hospital and clinics in Puerto Rico and Florida for the identification and recruitment of lung cancer patients and matched healthy controls. Once identified and consented, standardized procedures will be used for the administration of the surveys, collection of blood and sample processing at Moffitt Cancer Center; and (2) to assess the influence of genetic polymorphisms in candidate genes on lung cancer risk, we will compare allelic and genotype frequencies of TagSNPs covering selected candidate genes loci in 150 lung cancer patients and 150 matched controls of Puerto Rican origin. Combined analysis of the questionnaires and genetic studies will provide us with insight into single SNP loci and geneenvironmental interactions on the lung cancer phenotype. Understanding the contribution of genetic and environmental factors to lung cancer susceptibility in specific populations might provide insight into the mechanisms involved in lung cancer development.

This proposal combines the efforts of Dr. Gabrilovich, who will lead the experimental studies, and Dr. Antonia, who will supervise experiments related to clinical material. The ultimate goal is to develop a full-length SPORE proposal focused on translational study of TRAILR targeting in lung cancer. This future proposal will include abasic biology study of MDSC and the immune responses in tumor-bearing mice, and several clinical trials: one, a phase II trial in patients with NSCLC, aiming to investigate the effect of TRAILR2 on MDSC and non-specific immune responses; and the second, a larger follow-up trial to investigate the combination of TRAILR2 targeting with cancer vaccine. We believe that these studies will shed light on the molecular mechanisms underlying stemness of NSCLC cells. These studies will enable us to submit a RO1 grant in the very near future.

2. Sept 2008 - Aug 2010

Sept 2009 - Aug 2010
SPORE Developmental Projects

Developmental Research Projects (09/10 – Year 2)
The SPORE funded four developmental research projects as follows:

  • CHK1 as a predictor of synergy between HDAC inhibitors and chemotherapeutic drugs: Soner Altiok
  • Regulation of oncogene-induced imflammatory gene expression and lung tumorigenesis by NF-kappaB/IKKbeta: Amer Beg
  • nAChR mediated survival of non-small cell lung cancer cells: Srikumar Chellappan
  • Evaluation of Shp2 as a Novel Therapeutic Target in Lung Cancer: Jie Wu

Abstracts

CHK1 as a predictor of synergy between HDAC inhibitors and chemotherapeutic drugs: Soner Altiok

Histone deacetylase inhibitors (HDACi) represent a promising new treatment for cancer therapy. However, in clinical trials the effectiveness of HDACis was limited and no clear mechanism of resistance has been elucidated. Several preclinical reports have shown that HDAC inhibitors synergize with cytotoxic agents to induce apoptotic cell death in tumor cells. However, the molecular mechanisms of this synergistic interaction are not well understood. One of the major challenges in the development of HDACis is the definition of the clinically relevant endpoints to assess drug efficacy in patients’ tumors at the early stage of treatment to predict clinical outcome.

One validated target protein that regulates the G2 checkpoints is a kinase, CHK1. CHK1 is essential for cells to progress normally through an unperturbed cell division cycle and to arrest in response to checkpoint activation by DNA damaging agents. We have recently demonstrated that HDAC inhibition leads to degradation of CHK1 protein and, consequently, activation of CDC2/CDC25 mitotic regulators in non-small cell lung cancer cell (NSCLC) lines, and in clinical tumor samples. Furthermore, ex vivo experiments performed with primary NSCLC mouse xenografts developed in our laboratory showed that an HDAC inhibitor, LBH589, enhances the cytotoxic effects of gemcitabine, carboplatin and pemetrexed in tumors that exhibit CHK1 downregulation and CDC2 activation upon LBH589 treatment. Based on our preliminary data we hypothesize that HDACi-induced CHK1 downregulation plays a key role in response to HDACi treatment and in the enhancement of tumor sensitivity to chemotherapy by HDACis. Thus, the specific aim of this application is to analyze the potential role of CHK1 pathway inhibition in sensitivity to HDACi treatment and to predict synergy between HDACi and DNA damaging drug therapy in primary NSCLC mouse xenograft models.

The results of the proposed studies are expected to fill in gaps in our knowledge regarding the molecular mechanisms of resistance to HDACis and synergy between HDACi and cytotoxic agents. This information may provide a basis for the development of novel anti-cancer therapeutic strategies where validation of CHK1 as an early PD marker of HDACi therapy will have direct application for the enrichment of clinical trials and individualized cancer therapy.

Regulation of oncogene-induced imflammatory gene expression and lung tumorigenesis by NF-kappaB/IKKbeta: Amer Beg

Lung cancer is a leading cause of cancer-related death around the world for which the 5-year survival has remained unchanged over many decades. New targets and novel approaches will be required to substantially impact the efficacy of lung cancer therapy. The studies proposed here will investigate a newly discovered role for inflammatory cytokines in lung tumorigenesis. Specifically, we will investigate the role of the NF-kappaB/IKKbeta pathway in oncogene-induced inflammatory cytokine expression and lung tumor development. Two aims are proposed: Aim 1: Regulation of NF-kappaB/IKKbeta activation and inflammatory gene expression by oncogenic K-ras and EGFR. Aim 2: Lung tumor development in vivo by conditional K-ras expression.

nAChR mediated survival of non-small cell lung cancer cells: Srikumar Chellappan

Non-small cell lung cancer (NSCLC) demonstrates a strong etiologic association with smoking. It has been shown that physiological concentrations of nicotine could induce cell proliferation and angiogenesis in vitro and in vivo. Our lab had demonstrated that nicotine significantly promotes the progression and metastasis of tumors in mouse models of lung cancer, when nicotine was administered through intraperitoneal injections, or through over-the-counter transdermal patches. We had shown that nicotine can confer resistance to cellular apoptosis induced by chemotherapeutic agents like gemcitabine, taxols and cisplatin. These results suggested that exposure to nicotine, through nicotine supplements as well as smoking, could impair the beneficial effects of chemotherapeutic agents. Studies proposed in this DRP aim to understand the molecular mechanisms involved in nicotine-mediated resistance to apoptosis and will assess the role of PI3 kinase pathway and E2F1 transcription factor in the process. Further, the effect of nAChR agonist Varenidine/Varenicline (Chantix) on cell proliferation as well as apoptosis induced by chemotherapy drugs will be examined. In preliminary BrdU incorporation assays Chantix at 0.5?M and 1?M concentrations was as effective as 1?M nicotine in inducing the proliferation of A549 cells when compared to quiescent cells. We also carried out a TUNEL assay for apoptosis to see if Chantix can increase the survival of NSCLC cells. 0.1?M and 1?M Chantix decreased apoptosis comparable to 1?M nicotine (0.5-fold) in serum starved A549 cells.

Based on these preliminary results, we will continue to examine whether Chantix confers resistance to apoptosis induced by chemotherapeutic drugs in A549 cells.

We will also assess the contribution of PI3 Kinase pathway and E2F1 in mediating nicotine- or Chantix-mediated resistance to apoptosis. We believe that these studies will reveal the molecular mechanisms by which nicotine and other nAChR agonists confer resistance to apoptosis.

Evaluation of Shp2 as a Novel Therapeutic Target in Lung Cancer: Jie Wu

Increasing evidence suggests that cell signaling is coordinately regulated by both protein tyrosine kinases (PTKs) and protein tyrosine phosphatases (PTPs). Like PTKs, some of PTPs may be required for tumorigenesis and metastasis of human cancer. Shp2 is a non-receptor PTP encoded by the human PTPN11 gene. Gain-of-function Shp2 mutants have been linked to various hematologic malignancies. While the role of Shp2 in carcinoma is less clear, Shp2 is phosphorylated by receptor tyrosine kinases such as EGFR, MET, ROS, PDGFR, and ALK in lung cancer cells. To determine the effect of Shp2 phosphorylation, we created a phosphorylation mimicking mutant of Tyr62 (Y62D) and determined its PTP activity. Our data suggested that Tyr62 phosphorylation activated the Shp2 PTP activity. To begin evaluating Shp2 as a potential therapeutic target in lung carcinoma, we used doxycycline (dox)-inducible shRNAs to knockdown Shp2 in H292 and H358 non-small cell lung carcinoma cells as well as in non-transformed hTBE cells and in HEK293 cells. Our data showed that Shp2 knockdown preferentially inhibited proliferation of carcinoma cells. Analysis of H292 cells containing dox-inducible Shp2 shRNAs indicated that Shp2 knockdown prevented cell cycle progression, inhibited Erk1/2 and Src activation, reduced c-Myc, and increased p27 (CDKN1B) level. Significantly, Shp2 knockdown prevented tumor growth of H292 cells in nude mice. While H358 cells harbor a KRAS-G12C mutant, Shp2 inhibition also regulated c-Myc and p27 expression and prevented anchorage-independent growth of these cells. These results suggest that Shp2 is critically involved in malignant growth of lung cancer and a potential target for development of a novel PTP-targeted therapy in lung cancer.

Sept 2008 - Aug 2009
SPORE Developmental Projects

Developmental Research Projects (08/09 – Year 1)

The SPORE funded four developmental research projects as follows:

  • Novel mechanism of tumor escape in cancer: Dmitry Gabrilovich  
  • Role of Stat3 and Id transcription factors in Non-Small Cell Lung Cancer: Srikumar Chellappan and Eric Haura
  • Targeting the AKT Pathway in Lung Cancer: Jin Cheng and Gerold Bepler
  • BRCA1 status as a marker of clinical outcome in Lung Cancer: Alvaro Monteiro and Gerold Bepler

Abstracts

Novel mechanism of tumor escape in cancer: Dmitry Gabrilovich

Despite high promise the immunotherapeutic strategies that have been tested in clinical trials have not yet delivered tangible benefits to most patients. Numerous clinical trials have demonstrated successful generation of immune responses against tumor-specific antigens. However, these responses have not been associated with clinical responses in most patients. It has become increasingly clear that one of the major factors that limit the effectiveness of immunotherapy is the immunosuppressive tumor microenvironment, which prevents cytotoxic T cells (CTL) from recognizing and eliminating tumor. Recently, we and others have described an important role of myeloid derived suppressor cells (MDSC) derived reactive oxygen and nitrogen species in tumor evasion of immune mediated killing. Superoxide reacts with nitric oxide to form peroxynitrite, a highly reactive substance that modifies various proteins important for cell function. We have shown that peroxynitrite produced by MDSC can modify surface molecules on T cells that results in their inability to recognize and respond to peptide epitopes. Previous studies have demonstrated that lung cancer tissues contain high levels of nitrotyrosine, one of the hallmarks of peroxynitrite activity. We propose here that one of the major mechanisms of tumor escape in lung cancer is modification of MHC class I molecules expressed on tumor cells and/or tumor-specific peptides presented by those MHC class I molecules. These modifications are caused by peroxynitrite and ROS produced by myeloid and tumor cells. This would make CTLs specific for tumor antigen derived peptides unable to recognize and eliminate tumor cells. Thus, even if cancer immunotherapy results in the generation of potent antigen-specific CTLs or if these CTLs are generated in vitro and then adoptively transferred to patients, post-translational modification of MHC class I would negate any possible antitumor effect. Since the level of myeloid cell infiltration and peroxynitrite production varies from patient to patient this may account for variable results of patients’ treatment with adoptive transfer of T cells or vaccines.

Role of Stat3 and Id transcription factors in Non-Small Cell Lung Cancer: Srikumar Chellappan and Eric Haura

Non-small cell lung carcinoma is highly correlated with smoking and smokers constitute about 75% of NSCLC patients. Many tobacco-specific carcinogens present in cigarette smoke cause DNA damage, leading to the mutation of vital genes like Ras, p53 and Rb. In addition, many of these tobacco carcinogens as well as nicotine itself can promote cell proliferation and angiogenesis through the activation of nicotinic acetylcholine receptors. NSCLC in smokers and non-smokers are qualitatively different and have different molecular signatures; for example, cancers in non-smokers have EGFR mutations predominantly. Though NSCLC arises due to different but similar molecular events that lead to increased proliferation, tumor growth, angiogenesis and metastasis, we believe that there could be common elements in the signaling pathway that mediate the oncogenic process in smokers and non-smokers. We hypothesize the transcription factors STAT3 and Id1 play a major role in mediating the oncogenic process in NSCLC. Both these factors can be induced by nicotine and are known to correlate with a more advanced and progressed phenotype. Studies proposed in this application will examine how STAT3 and Id1 mediate the growth, progression and metastasis of NSCLC using a variety of in vitro and in vivo experiments. The molecular events mediating their angiogenic ans well as pro-metastatic properties will be elucidated. In addition, experiments are also proposed to examine how the levels of STAT3 and Id1 correlate with tumor stage and overall survival of NSCLC patients. We believe that these studies will shed light on novel mechanisms underlying the growth and progression of lung cancers.

Targeting the AKT Pathway in Lung Cancer: Jin Cheng and Gerold Bepler

The AKT pathway is activated in more than half of human lung cancers, and activation of AKT renders lung cancer cells resistant to conventional chemoradiotherapy and tyrosine kinase inhibitors (TKIs). Therefore, the AKT pathway represents a critical therapeutic target for improvement of lung cancer treatment. By screening of the NCI compound library, we have identified two AKT inhibitors, one of which (API-2/triciribine/TCN) has previously been used as a cytotoxic drug in clinic trials; however, without knowing its molecular target, and the other (API-1/AKT-SI1) is an uncharacterized compound in human cancer. Both API-1 and API-2 inhibited three members of the AKT family as well as constitutively active Akt including the naturally occurring activated AKT1-E17K mutation. We hypothesize that API-1 and API-2 can selectively inhibit AKT-dependant lung cancer cell growth and enhance the efficacy of conventional chemotherapy, radiation and small molecule TKIs in lung cancer. To test this, we propose to: 1, determine anti-tumor activity of the AKT inhibitors API-1 and API-2 in lung cancer with defined AKT status in vitro and in vivo, 2. examine the effects of AKT inhibitors on the AKT activation that is resulted from mTOR targeted therapy and acquired secondary alterations of EGFR in NSCLC and 3. determine if API-1 and API-2 sensitize high p-AKT lung cancer cells to apoptosis and growth arrest by chemotherapeutic agents and radiation as a means to overcome chemoradioresistance.

BRCA1 status as a marker of clinical outcome in Lung Cancer: Alvaro Monteiro and Gerold Bepler

The main agents used for chemotherapy for non-small cell lung cancer (NSCLC) today are nucleoside inhibitors and platinum analogs, combined with microtubule-interfering drugs such as the taxanes. However, many patients suffer the serious side effects without experiencing a significant disease response. Therefore, the identification of markers that can predict response to specific drugs is a top priority in lung cancer management.

The field has shown significant progress by the demonstration that EGFR mutation status may predict response to gefitinib and erlotinib. Recently we provided results indicating that the status of RRM1 and of ERCC1 can predict response to carboplatin and gemcitabine. Conspicuously absent is a marker to predict response to taxanes. Here we aim to fill this gap by testing the hypothesis that BRCA1 status correlates with response to taxanes in vitro and in the clinic, and can be developed as a predictor of drug response. 

 

 
 
 
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