Anthony Saleh - Academia.edu (original) (raw)

Papers by Anthony Saleh

Molecular Cancer Research

TNFα is a key mediator of immune and radiotherapy-induced cytotoxicity, but many cancers, includi... more TNFα is a key mediator of immune and radiotherapy-induced cytotoxicity, but many cancers, including head and neck squamous cell carcinomas (HNSCC), display TNF resistance due to activation of the canonical IKK–NF-κB/RELA pro-survival pathway. However, toxicities associated with direct targeting of the canonical pathway point to the need to identify mechanism(s) contributing to TNFα resistance and synthetic lethal targets to overcome such resistance in cancer cells. Here, RNAi screening for modulators of TNFα–NF-κB reporter activity and cell survival unexpectedly implicated the WEE1 and CDC2 G2–M checkpoint kinases. The IKKα/β-RELA and WEE1-CDC2 signaling pathways are activated by TNFα and form a complex in cell lines derived from both human papillomavirus (−) and (+) subtypes of HNSCC. WEE1 inhibitor AZD1775 reduced IKK/RELA phosphorylation and the expression of NF-κB–dependent pro-survival proteins Cyclin D1 and BCL2. Combination of TNFα and AZD1775 enhanced caspase-mediated apopto...

Cancer Research, 2009

Introduction: Radiotherapy can be complicated by debilitating normal tissue toxicity such as radi... more Introduction: Radiotherapy can be complicated by debilitating normal tissue toxicity such as radiation-induced fibrosis. We have previously shown that mir-21 and let-7b expression is altered by radiation in vitro . We hypothesize that these miRNA species may target proteins involved in radiation-induced fibrosis and therefore may serve as clinical targets to prevent morbidity. TGF\#946; is associated with induction of fibrosis, but has been reported to be down-regulated in long-term fibrosis. The aim of this study is to demonstrate that radiation alters the expression of miRNA species in vivo that target fibrosis-associated proteins such as TGF\#946;. Methods: The right hind leg of 9 week old CH3 mice was irradiated to 35Gy in a single fraction with the left leg serving as an unirradiated control. Mice were euthanized at 2hrs, 7 days, 14 days, 30 days or 150 days after irradiation. Fibrosis in the irradiated leg was determined by the amount of leg shortening compared to the unirradi...

Nutrition & Metabolism, 2020

Background Metastatic cancer is incurable and understanding the molecular underpinnings is crucia... more Background Metastatic cancer is incurable and understanding the molecular underpinnings is crucial to improving survival for our patients. The IGF-1/Akt signaling pathway is often impaired in cancer leading to its progression and metastases. Diet modification is known to alter the IGF-1/Akt pathway and affect the expression of microRNA involved in tumor initiation, growth and metastases. Liver metastases are one of the most common type of metastases in breast and colon cancer. In the present study, we looked at the effect of diet modification on the expression of microRNA in normal liver and liver with breast cancer metastases using in vivo model. Methodology 6-month-old C57BL/6 J mice were put on either an ad libitum (AL) diet, or 40% calorie restricted (CR) diet or were fasted for 24 h (FA) before sacrifice. MicroRNA array analysis, western blot and qRT-PCR were performed using liver tissue to compare the treatment groups. A breast cancer model was also used to study the changes i...

Cancer Research, 2020

Cytotoxic and genotoxic therapies are known to induce the prosurvival Inhibitor-kappaB kinase (IK... more Cytotoxic and genotoxic therapies are known to induce the prosurvival Inhibitor-kappaB kinase (IKK)-NF-κB/RELA pathway and DNA repair at the G2/M checkpoint, thereby promoting therapeutic resistance, but the coordination of these mechanisms is unclear. Our recent RNAi screen unexpectedly uncovered possible crosstalk between WEE1, a G2/M checkpoint kinase and the TNFα-IKK-NF-κB prosurvival pathway components. siRNAs targeting WEE1 and IKKs both inhibited TNFα-inducible NF-κB activity in a HNSCC reporter line, suggesting a mechanistic linkage. We investigated this potential connection further and studied the effect of WEE1 inhibition using AZD1775 in head and neck squamous cell carcinoma (HNSCC) cells and mouse xenograft models. Increased expression, phosphorylation and cellular localization of IKK-NF-κB and WEE1-CDC2 axis proteins co-occurred in a subset of HNSCC lines, and were inversely co-modulated by TNFα and WEE1 kinase inhibitor AZD1775. WEE1 co-immunoprecipitated with the IKKα...

Drug Metabolism and Pharmacokinetics, 2019

Molecular Carcinogenesis, 2018

Head and neck squamous cell carcinomas (HNSCC) preferentially spread to regional cervical tissues... more Head and neck squamous cell carcinomas (HNSCC) preferentially spread to regional cervical tissues and lymph nodes. Here, we hypothesized that lymphotoxin‐β (LTβ), receptor LTβR, and NF‐κB‐inducing kinase (NIK), promote the aberrant activation of alternative NF‐κB2/RELB pathway and genes, that enhance migration and invasion of HNSCC. Genomic and expression alterations of the alternative NF‐kB pathway were examined in 279 HNSCC tumors from The Cancer Genome Atlas (TCGA) and a panel of HNSCC lines. LTβR is amplified or overexpressed in HNSCC of the larynx or oral cavity, while LTβ, NIK, and RELB are overexpressed in cancers arising within lymphoid oropharyngeal and tonsillar sites. Similarly, subsets of HNSCC lines displayed overexpression of LTβR, NIK, and RELB proteins. Recombinant LTβ, and siRNA depletion of endogenous LTβR and NIK, modulated expression of LTβR, NIK, and nuclear translocation of NF‐κB2(p52)/RELB as well as functional NF‐κB promoter reporter activity. Treatment with ...

OncoImmunology, 2018

Intrinsic resistance to cytotoxic T-lymphocyte (CTL) killing limits responses to immune activatin... more Intrinsic resistance to cytotoxic T-lymphocyte (CTL) killing limits responses to immune activating anticancer therapies. Here, we established that activation of the G2/M cell cycle checkpoint results in tumor cell cycle pause and protection from granzyme B-induced cell death. This was reversed with WEE1 kinase inhibition, leading to enhanced CTL killing of antigen-positive tumor cells. Similarly, but at a later time point, cell cycle pause following TNFα exposure was reversed with WEE1 kinase inhibition, leading to CTL transmembrane TNFα-dependent induction of apoptosis and necroptosis in bystander antigennegative tumor cells. Results were reproducible in models of oral cavity carcinoma, melanoma and colon adenocarcinoma harboring variable Tp53 genomic alterations. WEE1 kinase inhibition sensitized tumors to PD-1 mAb immune checkpoint blockade in vivo, resulting in CD8 +-dependent rejection of established tumors harboring antigen-positive or mixed antigen-positive and negative tumor cells. Together, these data describe activation of the G2/M cell cycle checkpoint in response to early and late CTL products as a mechanism of resistance to CTL killing, and provide pre-clinical rationale for the clinical combination of agents that inhibit cell cycle checkpoints and activate anti-tumor immunity.

BMC cancer, Jan 2, 2017

Breast cancer is the most common invasive cancer among women. Currently, there are only a few mod... more Breast cancer is the most common invasive cancer among women. Currently, there are only a few models used for therapy selection, and they are often poor predictors of therapeutic response or take months to set up and assay. In this report, we introduce a microfluidic OrganoPlate® platform for extracellular matrix (ECM) embedded tumor culture under perfusion as an initial study designed to investigate the feasibility of adapting this technology for therapy selection. The triple negative breast cancer cell lines MDA-MB-453, MDA-MB-231 and HCC1937 were selected based on their different BRCA1 and P53 status, and were seeded in the platform. We evaluate seeding densities, ECM composition (Matrigel®, BME2rgf, collagen I) and biomechanical (perfusion vs static) conditions. We then exposed the cells to a series of anti-cancer drugs (paclitaxel, olaparib, cisplatin) and compared their responses to those in 2D cultures. Finally, we generated cisplatin dose responses in 3D cultures of breast c...

Cancer Research, 2017

As part of the Cancer Genome Atlas (TCGA) Network, our comprehensive genomic analysis of 279 head... more As part of the Cancer Genome Atlas (TCGA) Network, our comprehensive genomic analysis of 279 head and neck squamous cell carcinomas (HNSCCs) found frequent chromosomal copy number variation (CNV) and mutations of potential biologic and therapeutic importance. This underscored an urgent need to identify cell line models that harbor genomic alterations representative of HNSCC. We performed whole exome-DNA and transcriptome RNA sequencing on 15 human papillomavirus HPV(−) and 11 HPV(+) HNSCC cell lines. HNSCC lines harbored chromosome gains (3q, 5p, 7p, 8q, 11q) and losses (3p, 5q, 8p, 9p, 18q), consistent with those found in HNSCC tumors by TCGA and previous karyotype studies. Integrative genome-wide analysis of CNV with gene expression uncovered over 1500 genes that display significant correlation between CNV and gene expression in both TCGA tumors and cell lines. Ingenuity Pathway Analysis revealed multiple genes that converge on key pathways and functions deregulated in HNSCC, incl...

Cancer Research, 2016

Head and neck squamous cell carcinoma (HNSCC) is the most prevalent cancer affecting the upper ae... more Head and neck squamous cell carcinoma (HNSCC) is the most prevalent cancer affecting the upper aerodigestive tract, with an annual incidence of 600,000 patients and a five year survival of approximately 60% worldwide. Molecular mechanisms driving the development of HNSCC have recently begun to be discovered, with The Cancer Genome Atlas (TCGA) uncovering the genomic landscape of 279 cases of HNSCC. Alterations in cell death pathways were commonly found in the TCGA analysis, with ∼30% of samples harboring 11q13/22 amplifications and overexpression of genes encoding for Fas-associated death domain (FADD) and/or cellular Inhibitor of Apoptosis Proteins 1/2 (cIAP1/2). While overexpression of cIAP1 has been implicated in resistance to cytotoxic therapies, the role of FADD amplification as a target for therapy and in mechanisms of cell death is not well understood. Birinapant is a novel second mitochondria-derived activator of caspases (SMAC)-mimetic that targets and promotes degradation ...

Cancer research, Sep 28, 2016

Comparison of tumors from the Cancer Genome Atlas (TCGA) reveals that head and neck squamous cell... more Comparison of tumors from the Cancer Genome Atlas (TCGA) reveals that head and neck squamous cell carcinomas (HNSCC) harbor the most frequent genomic amplifications of Fas-associated death domain (FADD), with or without Baculovirus Inhibitor of Apoptosis repeat containing BIRC2 (cIAP1), affecting ~30% of patients in association with worse prognosis. Here, we identified HNSCC cell lines harboring FADD/BIRC2 amplifications and overexpression by exome sequencing, RT-PCR and Western blot. In vitro, FADD or BIRC2 siRNA knockdown inhibited HNSCC displaying amplification and increased expression of these genes, supporting their functional importance in promoting proliferation. Birinapant, a novel SMAC mimetic, sensitized multiple HNSCC lines to cell death by agonists TNFαalpha or TRAIL, and inhibited cIAP1>XIAP>IAP2. Combination of birinapant and TNFα induced sub-G0 DNA fragmentation in sensitive lines, and birinapant alone also induced significant G2/M cell cycle arrest and cell dea...

Cancer Research, 2015

Head and neck squamous cell carcinoma (HNSCC) is the 6th most common cancer worldwide with a 50-6... more Head and neck squamous cell carcinoma (HNSCC) is the 6th most common cancer worldwide with a 50-60% mortality rate. Deregulation of p53 family members in HNSCC occurs in over 90% of cases, preventing transcription of growth arrest and apoptosis genes. Conversely, members of the NF-κB/REL family are aberrantly activated in about ∼70% of cases, and drive expression of pro-proliferation, inflammation, angiogenesis, and therapeutic resistance genes. The function of different TP53 and NF-κB family members are inversely modulated within two major subsets of HNSCC, suggesting that common molecules and pathways coordinate this modulation. To test this hypothesis, we have developed HNSCC stable cell lines that report transcriptional activation of TP53 or NF-κB individually through a β-lactamase reporter. Whole genome siRNA screening in our NF-κB reporter cell line has been performed. Screening was performed by 56-hour knockdown with siRNAs, followed by stimulation with TNF-α. The results hav...

Cancer Research, 2014

Head and neck squamous cell carcinoma (HNSCC) is the 6th most common cancer, with an annual incid... more Head and neck squamous cell carcinoma (HNSCC) is the 6th most common cancer, with an annual incidence of over 600,000 new cases worldwide, and 50-60% mortality. Dysregulation of tumor suppressor TP53 in HNSCC occurs in over 90% of cases. We and others have observed that TP53 related family members p63 and p73 are also deregulated, together preventing transcription of growth arrest and apoptosis genes. Conversely, we have shown that members of the NF-κB/REL family of transcription factors are aberrantly activated, and drive expression of genes that promote cell proliferation, survival, inflammation, angiogenesis,and resistance in ∼70% of HNSCC cases. We have previously published that the function of different TP53 and NF-κB family members are inversely modulated within two major subsets of HNSCC, suggesting that common molecules and pathways coordinate this modulation and may serve as critical targets for therapy. To test this hypothesis, we have developed HNSCC stable cell lines tha...

Cancer Research, 2014

Our laboratory previously discovered that aberrant NF-κB signaling and activation plays a critica... more Our laboratory previously discovered that aberrant NF-κB signaling and activation plays a critical role in promoting the oncogenic progression of head and neck squamous cell carcinoma (HNSCC). The Cancer Genome Atlas (TCGA) project, which investigated 279 HNSCC tissue specimens, uncovered significant genomic alterations of key molecules involved in NF-κB and death signaling, including amplification of FADD and BIRC2/3, mutation of caspase-8, and altered RIPKs in HPV(-), or deletion of TRAF3 in HPV(+) HNSCC tissues. Using PRISM (Protein Interactions by Structural Matching), we modeled protein-protein interaction complexes three dimensionally and identified ∼30 proteins that potentially interact with these genetically altered molecules involved in NF-κB and death pathways. Among more than 20 major cancer types that TCGA project has investigated, we found that HNSCC ranks among the top cancers harboring alterations of these genes, which are present in more than 70% of HNSCC tumors. To ...

Cancer Research, 2014

Head and neck squamous cell carcinoma (HNSCC) is the sixth most common cancer worldwide, and stan... more Head and neck squamous cell carcinoma (HNSCC) is the sixth most common cancer worldwide, and standard treatment with surgery or chemoradiation has significant morbidity, with 5 year survival of <50%. Personalized targeted therapies with small molecule inhibitors have gained interest following identification of genetic drivers from The Cancer Genome Atlas (TCGA). TCGA data shows >60% of head and neck tumors harbor genomic alterations in the RTK-PI3K-mTOR and MAPK pathways important in cell growth and survival. In this study, we determined the effects of the novel PI3K-mTOR inhibitor PF-5212384 (PF-384) on molecular targets and HNSCC growth in preclinical models. PF-384 IC50s of 0.75nM-133nM were found in 12 HNSCC lines by XTT cell density assays and treatment resulted in increased sub-G0 cell death and G0/G1 phase blockade by DNA flow cytometry. PF-384 strongly inhibited direct targets of PI3K-mTOR, aberrant NF-kB transactivation and induced cytokines, but only partially inhibi...

Methods in molecular biology (Clifton, N.J.), 2014

Understanding the biological relevance and context of microRNA (miRNA) regulation of target mRNAs... more Understanding the biological relevance and context of microRNA (miRNA) regulation of target mRNAs is difficult to ascertain because an individual miRNA aids simultaneously in the regulation of hundreds of mRNAs in a cell. With the increasing availability of large public datasets that profile both mRNA and miRNA expression levels from the same samples, it is possible to apply robust statistical methods to identify global negative correlations in miRNA and target mRNA expression. Using a dataset from The Cancer Genome Atlas as a case study, we show how to use linear regression analysis followed by permutation-based false discovery rate to assign high statistical power to pair-wise negative correlations of miRNA and mRNA expression. Used in conjunction with available prediction tools or other target databases, a high confidence dataset of global miRNA-mRNA interactions can be generated. We also describe further methods to prioritize identified interactions by integrating with mutation,...

Journal of Bronchology & Interventional Pulmonology, 2010

The purpose of this article is to describe 3 patients each of whom developed a different form of ... more The purpose of this article is to describe 3 patients each of whom developed a different form of tracheobronchial aspergillosis. We describe our clinical experience with 3 patients who developed significant respiratory symptoms secondary to Aspergillus infection. All patients were followed closely until there was resolution of pulmonary problems or they succumbed to respiratory insufficiency. The first patient had asthma and her clinical and bronchoscopic findings were compatible with the diagnosis of mucoid impaction syndrome caused by Aspergillus. Response to therapy was excellent with complete recovery. The second and third patients had what we believe was tracheobronchial pseudomembranous aspergillosis. The precise reason for this complication in the second patient is unknown. The third patient was immunosuppressed and developed tracheobronchial aspergillosis. Despite aggressive therapy, both of these patients died. Diagnostic bronchoscopy was helpful in detecting the airway abnormalities and for obtaining respiratory specimens for culture. These cases show the diverse tracheobronchial manifestations of Aspergillus species. Diagnostic bronchoscopy was helpful in the diagnosis of airway involvement by aspergillus.

CHEST Journal, 2007

We report a case of a 65-year-old male patient who presented with gradually worsening dyspnea ove... more We report a case of a 65-year-old male patient who presented with gradually worsening dyspnea over 2 years. History was significant for smoking and the absence of any hazardous occupational exposure. The clinical findings at presentation included absent breath sounds and stony dullness on the left side, with tracheal deviation contralaterally and clubbing. A chest roentgenogram showed a left-sided opacity occupying almost the entire left hemithorax. A subsequent CT scan of the chest revealed an intrathoracic, extrapulmonary lesion producing a mediastinal shift. Surgical resection of the mass was performed, and pathology, along with immunohistochemical studies positive for CD34 and negative for epithelial markers, confirmed the diagnosis of solitary fibrous tumor of the pleura (SFTP). SFTP is a rare neoplasm, and diagnosis is often difficult. Suspicion of SFTP should arise in the setting of a patient presenting with a paucity of symptoms (except in the case of an accompanying paraneoplastic syndrome), the absence of exposure to asbestos, and a large mass with sharp margins and encapsulation seen on a chest radiograph.

Chest, 2008

he diagnostic workup for multiple calcified pul-T monary nodules can be a challenge for clinician... more he diagnostic workup for multiple calcified pul-T monary nodules can be a challenge for clinicians. The differential diagnosis of multiple calcified pulmonary nodules includes a broad spectrum of etiologies, the most important of which are neoplastic and infectious. A detailed medical history and radiographic description, with particular emphasis on the location and, to a lesser extent, the morphology, number, and size of nodules and other associated radiographic abnormalities usually help to narrow the differential diagnosis. This discussion focuses on the differential diagnosis and the workup of multiple calcified pulmonary nodules. CASE PRESENTATION A 73-year-old woman who was a lifelong nonsmoker was referred to the pulmonary clinic for mild dyspnea and dry cough of several months' duration. Her medical history was significant for systemic hypertension and osteoarthritis. She was receiving therapy with metoprolol, 25 mdd. She denied having any other symptoms, including chest pain, weight loss, hemoptysis, fevers, chills, and night sweats. The findings of her physical examination were unremarkable. A chest radiograph with a posteroanterior view (Fig 1) showed multiple ill-defined nodules of varying sizes (range, 0.5 to 3.0 cm) in both lung fields. A CT scan of the chest

Chest, 2009

INTRODUCTION: There have been many reports describing the reversibility of heart failure caused b... more INTRODUCTION: There have been many reports describing the reversibility of heart failure caused by thyrotoxicosis. The prompt manner in which the heart function normalized in the setting of thyroid storm merits discussion.

Molecular Cancer Research

TNFα is a key mediator of immune and radiotherapy-induced cytotoxicity, but many cancers, includi... more TNFα is a key mediator of immune and radiotherapy-induced cytotoxicity, but many cancers, including head and neck squamous cell carcinomas (HNSCC), display TNF resistance due to activation of the canonical IKK–NF-κB/RELA pro-survival pathway. However, toxicities associated with direct targeting of the canonical pathway point to the need to identify mechanism(s) contributing to TNFα resistance and synthetic lethal targets to overcome such resistance in cancer cells. Here, RNAi screening for modulators of TNFα–NF-κB reporter activity and cell survival unexpectedly implicated the WEE1 and CDC2 G2–M checkpoint kinases. The IKKα/β-RELA and WEE1-CDC2 signaling pathways are activated by TNFα and form a complex in cell lines derived from both human papillomavirus (−) and (+) subtypes of HNSCC. WEE1 inhibitor AZD1775 reduced IKK/RELA phosphorylation and the expression of NF-κB–dependent pro-survival proteins Cyclin D1 and BCL2. Combination of TNFα and AZD1775 enhanced caspase-mediated apopto...

Cancer Research, 2009

Introduction: Radiotherapy can be complicated by debilitating normal tissue toxicity such as radi... more Introduction: Radiotherapy can be complicated by debilitating normal tissue toxicity such as radiation-induced fibrosis. We have previously shown that mir-21 and let-7b expression is altered by radiation in vitro . We hypothesize that these miRNA species may target proteins involved in radiation-induced fibrosis and therefore may serve as clinical targets to prevent morbidity. TGF\#946; is associated with induction of fibrosis, but has been reported to be down-regulated in long-term fibrosis. The aim of this study is to demonstrate that radiation alters the expression of miRNA species in vivo that target fibrosis-associated proteins such as TGF\#946;. Methods: The right hind leg of 9 week old CH3 mice was irradiated to 35Gy in a single fraction with the left leg serving as an unirradiated control. Mice were euthanized at 2hrs, 7 days, 14 days, 30 days or 150 days after irradiation. Fibrosis in the irradiated leg was determined by the amount of leg shortening compared to the unirradi...

Nutrition & Metabolism, 2020

Background Metastatic cancer is incurable and understanding the molecular underpinnings is crucia... more Background Metastatic cancer is incurable and understanding the molecular underpinnings is crucial to improving survival for our patients. The IGF-1/Akt signaling pathway is often impaired in cancer leading to its progression and metastases. Diet modification is known to alter the IGF-1/Akt pathway and affect the expression of microRNA involved in tumor initiation, growth and metastases. Liver metastases are one of the most common type of metastases in breast and colon cancer. In the present study, we looked at the effect of diet modification on the expression of microRNA in normal liver and liver with breast cancer metastases using in vivo model. Methodology 6-month-old C57BL/6 J mice were put on either an ad libitum (AL) diet, or 40% calorie restricted (CR) diet or were fasted for 24 h (FA) before sacrifice. MicroRNA array analysis, western blot and qRT-PCR were performed using liver tissue to compare the treatment groups. A breast cancer model was also used to study the changes i...

Cancer Research, 2020

Cytotoxic and genotoxic therapies are known to induce the prosurvival Inhibitor-kappaB kinase (IK... more Cytotoxic and genotoxic therapies are known to induce the prosurvival Inhibitor-kappaB kinase (IKK)-NF-κB/RELA pathway and DNA repair at the G2/M checkpoint, thereby promoting therapeutic resistance, but the coordination of these mechanisms is unclear. Our recent RNAi screen unexpectedly uncovered possible crosstalk between WEE1, a G2/M checkpoint kinase and the TNFα-IKK-NF-κB prosurvival pathway components. siRNAs targeting WEE1 and IKKs both inhibited TNFα-inducible NF-κB activity in a HNSCC reporter line, suggesting a mechanistic linkage. We investigated this potential connection further and studied the effect of WEE1 inhibition using AZD1775 in head and neck squamous cell carcinoma (HNSCC) cells and mouse xenograft models. Increased expression, phosphorylation and cellular localization of IKK-NF-κB and WEE1-CDC2 axis proteins co-occurred in a subset of HNSCC lines, and were inversely co-modulated by TNFα and WEE1 kinase inhibitor AZD1775. WEE1 co-immunoprecipitated with the IKKα...

Drug Metabolism and Pharmacokinetics, 2019

Molecular Carcinogenesis, 2018

Head and neck squamous cell carcinomas (HNSCC) preferentially spread to regional cervical tissues... more Head and neck squamous cell carcinomas (HNSCC) preferentially spread to regional cervical tissues and lymph nodes. Here, we hypothesized that lymphotoxin‐β (LTβ), receptor LTβR, and NF‐κB‐inducing kinase (NIK), promote the aberrant activation of alternative NF‐κB2/RELB pathway and genes, that enhance migration and invasion of HNSCC. Genomic and expression alterations of the alternative NF‐kB pathway were examined in 279 HNSCC tumors from The Cancer Genome Atlas (TCGA) and a panel of HNSCC lines. LTβR is amplified or overexpressed in HNSCC of the larynx or oral cavity, while LTβ, NIK, and RELB are overexpressed in cancers arising within lymphoid oropharyngeal and tonsillar sites. Similarly, subsets of HNSCC lines displayed overexpression of LTβR, NIK, and RELB proteins. Recombinant LTβ, and siRNA depletion of endogenous LTβR and NIK, modulated expression of LTβR, NIK, and nuclear translocation of NF‐κB2(p52)/RELB as well as functional NF‐κB promoter reporter activity. Treatment with ...

OncoImmunology, 2018

Intrinsic resistance to cytotoxic T-lymphocyte (CTL) killing limits responses to immune activatin... more Intrinsic resistance to cytotoxic T-lymphocyte (CTL) killing limits responses to immune activating anticancer therapies. Here, we established that activation of the G2/M cell cycle checkpoint results in tumor cell cycle pause and protection from granzyme B-induced cell death. This was reversed with WEE1 kinase inhibition, leading to enhanced CTL killing of antigen-positive tumor cells. Similarly, but at a later time point, cell cycle pause following TNFα exposure was reversed with WEE1 kinase inhibition, leading to CTL transmembrane TNFα-dependent induction of apoptosis and necroptosis in bystander antigennegative tumor cells. Results were reproducible in models of oral cavity carcinoma, melanoma and colon adenocarcinoma harboring variable Tp53 genomic alterations. WEE1 kinase inhibition sensitized tumors to PD-1 mAb immune checkpoint blockade in vivo, resulting in CD8 +-dependent rejection of established tumors harboring antigen-positive or mixed antigen-positive and negative tumor cells. Together, these data describe activation of the G2/M cell cycle checkpoint in response to early and late CTL products as a mechanism of resistance to CTL killing, and provide pre-clinical rationale for the clinical combination of agents that inhibit cell cycle checkpoints and activate anti-tumor immunity.

BMC cancer, Jan 2, 2017

Breast cancer is the most common invasive cancer among women. Currently, there are only a few mod... more Breast cancer is the most common invasive cancer among women. Currently, there are only a few models used for therapy selection, and they are often poor predictors of therapeutic response or take months to set up and assay. In this report, we introduce a microfluidic OrganoPlate® platform for extracellular matrix (ECM) embedded tumor culture under perfusion as an initial study designed to investigate the feasibility of adapting this technology for therapy selection. The triple negative breast cancer cell lines MDA-MB-453, MDA-MB-231 and HCC1937 were selected based on their different BRCA1 and P53 status, and were seeded in the platform. We evaluate seeding densities, ECM composition (Matrigel®, BME2rgf, collagen I) and biomechanical (perfusion vs static) conditions. We then exposed the cells to a series of anti-cancer drugs (paclitaxel, olaparib, cisplatin) and compared their responses to those in 2D cultures. Finally, we generated cisplatin dose responses in 3D cultures of breast c...

Cancer Research, 2017

As part of the Cancer Genome Atlas (TCGA) Network, our comprehensive genomic analysis of 279 head... more As part of the Cancer Genome Atlas (TCGA) Network, our comprehensive genomic analysis of 279 head and neck squamous cell carcinomas (HNSCCs) found frequent chromosomal copy number variation (CNV) and mutations of potential biologic and therapeutic importance. This underscored an urgent need to identify cell line models that harbor genomic alterations representative of HNSCC. We performed whole exome-DNA and transcriptome RNA sequencing on 15 human papillomavirus HPV(−) and 11 HPV(+) HNSCC cell lines. HNSCC lines harbored chromosome gains (3q, 5p, 7p, 8q, 11q) and losses (3p, 5q, 8p, 9p, 18q), consistent with those found in HNSCC tumors by TCGA and previous karyotype studies. Integrative genome-wide analysis of CNV with gene expression uncovered over 1500 genes that display significant correlation between CNV and gene expression in both TCGA tumors and cell lines. Ingenuity Pathway Analysis revealed multiple genes that converge on key pathways and functions deregulated in HNSCC, incl...

Cancer Research, 2016

Head and neck squamous cell carcinoma (HNSCC) is the most prevalent cancer affecting the upper ae... more Head and neck squamous cell carcinoma (HNSCC) is the most prevalent cancer affecting the upper aerodigestive tract, with an annual incidence of 600,000 patients and a five year survival of approximately 60% worldwide. Molecular mechanisms driving the development of HNSCC have recently begun to be discovered, with The Cancer Genome Atlas (TCGA) uncovering the genomic landscape of 279 cases of HNSCC. Alterations in cell death pathways were commonly found in the TCGA analysis, with ∼30% of samples harboring 11q13/22 amplifications and overexpression of genes encoding for Fas-associated death domain (FADD) and/or cellular Inhibitor of Apoptosis Proteins 1/2 (cIAP1/2). While overexpression of cIAP1 has been implicated in resistance to cytotoxic therapies, the role of FADD amplification as a target for therapy and in mechanisms of cell death is not well understood. Birinapant is a novel second mitochondria-derived activator of caspases (SMAC)-mimetic that targets and promotes degradation ...

Cancer research, Sep 28, 2016

Comparison of tumors from the Cancer Genome Atlas (TCGA) reveals that head and neck squamous cell... more Comparison of tumors from the Cancer Genome Atlas (TCGA) reveals that head and neck squamous cell carcinomas (HNSCC) harbor the most frequent genomic amplifications of Fas-associated death domain (FADD), with or without Baculovirus Inhibitor of Apoptosis repeat containing BIRC2 (cIAP1), affecting ~30% of patients in association with worse prognosis. Here, we identified HNSCC cell lines harboring FADD/BIRC2 amplifications and overexpression by exome sequencing, RT-PCR and Western blot. In vitro, FADD or BIRC2 siRNA knockdown inhibited HNSCC displaying amplification and increased expression of these genes, supporting their functional importance in promoting proliferation. Birinapant, a novel SMAC mimetic, sensitized multiple HNSCC lines to cell death by agonists TNFαalpha or TRAIL, and inhibited cIAP1>XIAP>IAP2. Combination of birinapant and TNFα induced sub-G0 DNA fragmentation in sensitive lines, and birinapant alone also induced significant G2/M cell cycle arrest and cell dea...

Cancer Research, 2015

Head and neck squamous cell carcinoma (HNSCC) is the 6th most common cancer worldwide with a 50-6... more Head and neck squamous cell carcinoma (HNSCC) is the 6th most common cancer worldwide with a 50-60% mortality rate. Deregulation of p53 family members in HNSCC occurs in over 90% of cases, preventing transcription of growth arrest and apoptosis genes. Conversely, members of the NF-κB/REL family are aberrantly activated in about ∼70% of cases, and drive expression of pro-proliferation, inflammation, angiogenesis, and therapeutic resistance genes. The function of different TP53 and NF-κB family members are inversely modulated within two major subsets of HNSCC, suggesting that common molecules and pathways coordinate this modulation. To test this hypothesis, we have developed HNSCC stable cell lines that report transcriptional activation of TP53 or NF-κB individually through a β-lactamase reporter. Whole genome siRNA screening in our NF-κB reporter cell line has been performed. Screening was performed by 56-hour knockdown with siRNAs, followed by stimulation with TNF-α. The results hav...

Cancer Research, 2014

Head and neck squamous cell carcinoma (HNSCC) is the 6th most common cancer, with an annual incid... more Head and neck squamous cell carcinoma (HNSCC) is the 6th most common cancer, with an annual incidence of over 600,000 new cases worldwide, and 50-60% mortality. Dysregulation of tumor suppressor TP53 in HNSCC occurs in over 90% of cases. We and others have observed that TP53 related family members p63 and p73 are also deregulated, together preventing transcription of growth arrest and apoptosis genes. Conversely, we have shown that members of the NF-κB/REL family of transcription factors are aberrantly activated, and drive expression of genes that promote cell proliferation, survival, inflammation, angiogenesis,and resistance in ∼70% of HNSCC cases. We have previously published that the function of different TP53 and NF-κB family members are inversely modulated within two major subsets of HNSCC, suggesting that common molecules and pathways coordinate this modulation and may serve as critical targets for therapy. To test this hypothesis, we have developed HNSCC stable cell lines tha...

Cancer Research, 2014

Our laboratory previously discovered that aberrant NF-κB signaling and activation plays a critica... more Our laboratory previously discovered that aberrant NF-κB signaling and activation plays a critical role in promoting the oncogenic progression of head and neck squamous cell carcinoma (HNSCC). The Cancer Genome Atlas (TCGA) project, which investigated 279 HNSCC tissue specimens, uncovered significant genomic alterations of key molecules involved in NF-κB and death signaling, including amplification of FADD and BIRC2/3, mutation of caspase-8, and altered RIPKs in HPV(-), or deletion of TRAF3 in HPV(+) HNSCC tissues. Using PRISM (Protein Interactions by Structural Matching), we modeled protein-protein interaction complexes three dimensionally and identified ∼30 proteins that potentially interact with these genetically altered molecules involved in NF-κB and death pathways. Among more than 20 major cancer types that TCGA project has investigated, we found that HNSCC ranks among the top cancers harboring alterations of these genes, which are present in more than 70% of HNSCC tumors. To ...

Cancer Research, 2014

Head and neck squamous cell carcinoma (HNSCC) is the sixth most common cancer worldwide, and stan... more Head and neck squamous cell carcinoma (HNSCC) is the sixth most common cancer worldwide, and standard treatment with surgery or chemoradiation has significant morbidity, with 5 year survival of <50%. Personalized targeted therapies with small molecule inhibitors have gained interest following identification of genetic drivers from The Cancer Genome Atlas (TCGA). TCGA data shows >60% of head and neck tumors harbor genomic alterations in the RTK-PI3K-mTOR and MAPK pathways important in cell growth and survival. In this study, we determined the effects of the novel PI3K-mTOR inhibitor PF-5212384 (PF-384) on molecular targets and HNSCC growth in preclinical models. PF-384 IC50s of 0.75nM-133nM were found in 12 HNSCC lines by XTT cell density assays and treatment resulted in increased sub-G0 cell death and G0/G1 phase blockade by DNA flow cytometry. PF-384 strongly inhibited direct targets of PI3K-mTOR, aberrant NF-kB transactivation and induced cytokines, but only partially inhibi...

Methods in molecular biology (Clifton, N.J.), 2014

Understanding the biological relevance and context of microRNA (miRNA) regulation of target mRNAs... more Understanding the biological relevance and context of microRNA (miRNA) regulation of target mRNAs is difficult to ascertain because an individual miRNA aids simultaneously in the regulation of hundreds of mRNAs in a cell. With the increasing availability of large public datasets that profile both mRNA and miRNA expression levels from the same samples, it is possible to apply robust statistical methods to identify global negative correlations in miRNA and target mRNA expression. Using a dataset from The Cancer Genome Atlas as a case study, we show how to use linear regression analysis followed by permutation-based false discovery rate to assign high statistical power to pair-wise negative correlations of miRNA and mRNA expression. Used in conjunction with available prediction tools or other target databases, a high confidence dataset of global miRNA-mRNA interactions can be generated. We also describe further methods to prioritize identified interactions by integrating with mutation,...

Journal of Bronchology & Interventional Pulmonology, 2010

The purpose of this article is to describe 3 patients each of whom developed a different form of ... more The purpose of this article is to describe 3 patients each of whom developed a different form of tracheobronchial aspergillosis. We describe our clinical experience with 3 patients who developed significant respiratory symptoms secondary to Aspergillus infection. All patients were followed closely until there was resolution of pulmonary problems or they succumbed to respiratory insufficiency. The first patient had asthma and her clinical and bronchoscopic findings were compatible with the diagnosis of mucoid impaction syndrome caused by Aspergillus. Response to therapy was excellent with complete recovery. The second and third patients had what we believe was tracheobronchial pseudomembranous aspergillosis. The precise reason for this complication in the second patient is unknown. The third patient was immunosuppressed and developed tracheobronchial aspergillosis. Despite aggressive therapy, both of these patients died. Diagnostic bronchoscopy was helpful in detecting the airway abnormalities and for obtaining respiratory specimens for culture. These cases show the diverse tracheobronchial manifestations of Aspergillus species. Diagnostic bronchoscopy was helpful in the diagnosis of airway involvement by aspergillus.

CHEST Journal, 2007

We report a case of a 65-year-old male patient who presented with gradually worsening dyspnea ove... more We report a case of a 65-year-old male patient who presented with gradually worsening dyspnea over 2 years. History was significant for smoking and the absence of any hazardous occupational exposure. The clinical findings at presentation included absent breath sounds and stony dullness on the left side, with tracheal deviation contralaterally and clubbing. A chest roentgenogram showed a left-sided opacity occupying almost the entire left hemithorax. A subsequent CT scan of the chest revealed an intrathoracic, extrapulmonary lesion producing a mediastinal shift. Surgical resection of the mass was performed, and pathology, along with immunohistochemical studies positive for CD34 and negative for epithelial markers, confirmed the diagnosis of solitary fibrous tumor of the pleura (SFTP). SFTP is a rare neoplasm, and diagnosis is often difficult. Suspicion of SFTP should arise in the setting of a patient presenting with a paucity of symptoms (except in the case of an accompanying paraneoplastic syndrome), the absence of exposure to asbestos, and a large mass with sharp margins and encapsulation seen on a chest radiograph.

Chest, 2008

he diagnostic workup for multiple calcified pul-T monary nodules can be a challenge for clinician... more he diagnostic workup for multiple calcified pul-T monary nodules can be a challenge for clinicians. The differential diagnosis of multiple calcified pulmonary nodules includes a broad spectrum of etiologies, the most important of which are neoplastic and infectious. A detailed medical history and radiographic description, with particular emphasis on the location and, to a lesser extent, the morphology, number, and size of nodules and other associated radiographic abnormalities usually help to narrow the differential diagnosis. This discussion focuses on the differential diagnosis and the workup of multiple calcified pulmonary nodules. CASE PRESENTATION A 73-year-old woman who was a lifelong nonsmoker was referred to the pulmonary clinic for mild dyspnea and dry cough of several months' duration. Her medical history was significant for systemic hypertension and osteoarthritis. She was receiving therapy with metoprolol, 25 mdd. She denied having any other symptoms, including chest pain, weight loss, hemoptysis, fevers, chills, and night sweats. The findings of her physical examination were unremarkable. A chest radiograph with a posteroanterior view (Fig 1) showed multiple ill-defined nodules of varying sizes (range, 0.5 to 3.0 cm) in both lung fields. A CT scan of the chest

Chest, 2009

INTRODUCTION: There have been many reports describing the reversibility of heart failure caused b... more INTRODUCTION: There have been many reports describing the reversibility of heart failure caused by thyrotoxicosis. The prompt manner in which the heart function normalized in the setting of thyroid storm merits discussion.