Junaid Ansari

Epigenetics in non-small cell lung cancer: from basics to therapeutics

Lung cancer remains the number one cause of cancer-related deaths worldwide with 221,200 estimated new cases and 158,040 estimated deaths in 2015. Approximately 80% of cases are non-small cell lung cancer (NSCLC). The diagnosis is usually made at an advanced stage where the prognosis is poor and therapeutic options are limited. The evolution of lung cancer is a multistep process involving genetic, epigenetic, and environmental factor interactions that result in the dysregulation of key oncogenes and tumor suppressor genes, culminating in activation of cancer-related signaling pathways. The past decade has witnessed the discovery of multiple molecular aberrations that drive lung cancer growth, among which are epidermal growth factor receptor (EGFR) mutations and translocations involving the anaplastic lymphoma kinase (ALK) gene. This has translated into therapeutic agent developments that target these molecular alterations. The absence of targetable mutations in 50% of NSCLC cases and targeted therapy resistance development underscores the importance for developing alternative therapeutic strategies for treating lung cancer. Among these strategies, pharmacologic modulation of the epigenome has been used to treat lung cancer. Epigenetics approaches may circumvent the problem of tumor heterogeneity by affecting the expression of multiple tumor suppression genes (TSGs), halting tumor growth and survival. Moreover, it may be effective for tumors that are not driven by currently recognized druggable mutations. This review summarizes the molecular pathology of lung cancer epigenetic aberrations and discusses current efforts to target the epigenome with different pharmacological approaches. Our main focus will be on hypomethylating agents, histone deacetylase (HDAC) inhibitors, microRNA modulations, and the role of novel epigenetic biomarkers. Last, we will address the challenges that face this old-new strategy in treating lung cancer.

Histiocytic sarcoma as a secondary malignancy: pathobiology, diagnosis, and treatment

Histiocytic sarcoma (HS) is an extremely rare non‐Langerhans cell disorder with an aggressive course and limited treatment options. Recent advances in molecular/genetic sequencing have suggested a common clonal origin between various hematolymphoid disorders and cases of secondary HS. Deriving conclusions from previously reported cases of HS arising secondarily to certain hematolymphoid disorders, here we have tried to provide insight into the mechanisms influencing this evolution. We also discuss a clinical case of a 72‐year‐old man with a diagnosis of chronic myeloid leukemia (CML), presenting subsequently with a heterogeneous liver mass positive with a diagnosis of HS. The liver mass showed a retained BCR‐ABL1 translocation suggesting clonality between the CML and HS. As seen in our case and other reported cases of HS derived secondarily, the concurrent expression of immunoglobulin heavy (IGH)‐/light‐chain rearrangements or cytogenetic markers common to the primary malignancy suggests an evolutionary mechanism involving lineage switching that could potentially be influenced by genetic or epigenetic cues which may occur at the level of a progenitor or the malignant cell itself.

Therapeutic Potential of Annexin A1 in Ischemia Reperfusion Injury

Cardiovascular disease (CVD) continues to be the leading cause of death in the world. Increased inflammation and an enhanced thrombotic milieu represent two major complications of CVD, which can culminate into an ischemic event. Treatment for these life-threatening complications remains reperfusion and restoration of blood flow. However, reperfusion strategies may result in ischemia–reperfusion injury (I/RI) secondary to various cardiovascular pathologies, including myocardial infarction and stroke, by furthering the inflammatory and thrombotic responses and delivering inflammatory mediators to the affected tissue. Annexin A1 (AnxA1) and its mimetic peptides are endogenous anti-inflammatory and pro-resolving mediators, known to have significant effects in resolving inflammation in a variety of disease models. Mounting evidence suggests that AnxA1, which interacts with the formyl peptide receptor (FPR) family, may have a significant role in mitigating I/RI associated complications. In this review article, we focus on how AnxA1 plays a protective role in the I/R based vascular pathologies.

Sickle cell disease: a malady beyond a hemoglobin defect in cerebrovascular disease

ABSTRACT Introduction: Sickle cell disease (SCD) is a devastating monogenic disorder that presents as a multisystem illness and affects approximately 100,000 individuals in the United States alone. SCD management largely focuses on primary prevention, symptomatic treatment and targeting of hemoglobin polymerization and red blood cell sickling. Areas covered: This review will discuss the progress of SCD over the last few decades, highlighting some of the clinical (mainly cerebrovascular) and psychosocial challenges of SCD in the United States. In addition, focus will also be made on the evolving science and management of this inherited disease. Expert commentary: Until recently hydroxyurea (HU) has been the only FDA approved therapy for SCD. However, advancing understanding of SCD pathophysiology has led to multiple clinical trials targeting SCD related thrombo-inflammation, abnormal endothelial biology, increased oxidant stress and sickle cell mutation. Yet, despite advancing understanding, available therapies are limited. SCD also imposes great psychosocial challenges for the individual and the affected community, which has previously been under-recognized. This has created a pressing need for complementary adjuvant therapies with repurposed and novel drugs, in addition to the establishment of comprehensive clinics focusing on both the medical treatment and the psychosocial issues associated with SCD.

Hydrogen Sulfide and Hydrogen Sulfide-Synthesizing Enzymes Are Altered in a Case of Oral Adenoid Cystic Carcinoma

Adenoid cystic carcinomas (ACC) constitute 1% of all head and neck malignancies and are very rare in the oral cavity. With < 60 oral ACCs described, their pathobiology is incompletely understood. Here, we report a case of oral cavity ACC in a 54-year-old woman. Since recent studies have demonstrated that several human tumors overexpress the hydrogen sulfide (H2S)-synthesizing enzymes cystathionine-β-synthase (CBS), cystathionine γ-lyase (CSE), and 3-mercaptopyruvate sulfurtransferase (3-MST), and also show dysregulated H2S levels, we examined these biomarkers in the oral ACC and compared the results to those of adjacent benign oral epithelium. Western blotting was used to compare the protein expression of CBS, CSE, 3-MST, nicotinamide phosphoribosyl transferase, and mitoNEET in ACC and adjacent benign oral mucosae. High-performance liquid chromatography was used to quantify the differences in tissue H2S concentrations between the two biopsy types. We found that all the proteins examined here were increased in the ACC compared to adjacent benign oral mucosae. Interestingly, H2S concentrations were decreased approximately 30% in ACC compared to benign mucosae. Thus, in one example of this rare tumor type, the enzymes that synthesize H2S are increased, while tissue H2S levels are lower than those found in adjacent benign oral mucosae. Although limited to a single rare tumor type, to our knowledge this is the second time H2S concentrations have been directly quantified inside a human tumor. Last, our results may indicate that alterations in H2S synthesis and metabolism may be important in the pathobiology of ACC.

Non-celiac gluten sensitivity: All wheat attack is not celiac

Currently, 1% of the United States population holds a diagnosis for celiac disease (CD), however, a more recently recognized and possibly related condition, “non-celiac gluten sensitivity” (NCGS) has been suggested to affect up to 6% of the United States public. While reliable clinical tests for CD exist, diagnosing individuals affected by NCGS is still complicated by the lack of reliable biomarkers and reliance upon a broad set of intestinal and extra intestinal symptoms possibly provoked by gluten. NCGS has been proposed to exhibit an innate immune response activated by gluten and several other wheat proteins. At present, an enormous food industry has developed to supply gluten-free products (GFP) with GFP sales in 2014 approaching

Abstract 4816: Potent inhibition of the cell proliferation and induction of apoptosis in lymphoma cells by the anthelminthic drug niclosamide: in vitro data

1 billion, with estimations projecting sales to reach

Increased Nicotinamide Phosphoribosyltransferase in Rhabdomyosarcomas and Leiomyosarcomas Compared to Skeletal and Smooth Muscle Tissue

2 billion in the year 2020. The enormous demand for GFP also reflects a popular misconception among consumers that gluten avoidance is part of a healthy lifestyle choice. Features of NCGS and other gluten related disorders (e.g., irritable bowel syndrome) call for a review of current distinctive diagnostic criteria that distinguish each, and identification of biomarkers selective or specific for NCGS. The aim of this paper is to review our current understanding of NCGS, highlighting the remaining challenges and questions which may improve its diagnosis and treatment.

336 Increased Nicotinamide Phosphoribosyltransferase and Cystathionine B-Synthase in Renal Oncocytomas, Renal Transitional Cell Carcinoma, and Renal Cell Carcinoma

Background: Niclosamide, an anthelminthic drug, has demonstrated anti-cancer potential in variety of malignancies. However only a limited number of studies have been performed in lymphoma models, therefore we hypothesized that niclosamide may also have anti-cancer potential on B-cell lymphomas. Materials and Methods: Established B lymphoma cell lines were exposed to different concentrations of niclosamide and IC50 was calculated using GraphPad Prism 6.0 software. Cell viability and proliferation were assessed by CellTiter-Blue and trypan blue exclusion assays. Apoptosis was assessed by flow cytometry following Annexin-V/ propidium iodide staining. Gene expression changes were studied using GeneChip Human Transcriptome Array 2.0. Colony forming assays were performed in methylcellulose. Ultrastructural cellular changes were studied with electron microscopy. Peripheral blood mononuclear cells (PBMCs) from individuals without active cancer and from patients with different hematologic disorders, were also exposed with niclosamide. Results: Treatment with niclosamide resulted in time-and dose- dependent apoptosis, cytotoxicity and inhibition of proliferation in different lymphoma cell lines including vincristine-refractory cell line. The IC50 of lymphoma cells lines is as follows: Daudi: 0.33 iM; HBL-2: 0.57 iM; KOPN-8: 0.72 iM; Ramos: 0.53 iM and SU-DHL4-VR: 0.45 iM. Niclosamide also inhibited clonal growth in semi-solid media. Gene expression changes were studied in Daudi and KOPN-8 cells treated with 2.5 iM Niclosamide for 3 and 6 hours. 96 genes were consistently overexpressed, 59 down-regulated. 10 genes involved in the tumor necrosis factor (TNF) pathway and 10 genes involving the DNA damage pathway were overexpressed. 13 out of the 59 down-regulated genes were involved in mitochondrial function. Electron microscopy showed that filopodia increased and lipid vacuoles developed whereas mitochondria were less numerous in KOPN-8 cells. The viability of PBMCs from 8 individuals without lymphoma was unchanged when incubated with niclosamide, whereas niclosamide showed significant cytotoxicity in a patient with mantle cell lymphoma (MCL). Conclusion: Niclosamide effectively inhibits the proliferation of B lymphoma cell lines, including vincristine-refractory lymphoma cells, and induces apoptosis at concentrations non-toxic to PBMCs. Interestingly, niclosamide exhibited cytotoxic activity against MCL cells - a finding worth testing further in this difficult-to-treat disease. The mechanism of action of Niclosamide may involve the TNF receptor pathway, mitochondrial function and DNA damage response pathway. We plan to elucidate further specific mechanism(s) of action, and evaluate synergistic effects with other antineoplastic agents, and perform in vivo studies. Citation Format: Junaid Ansari, Hazem El-Osta, Paula Polk, Jeffrey J. Aufman, Guillermo A. Herrera, James Cardelli, Rodney E. Shackelford, Glenn M. Mills, Magdalena L. Circu, Felicity N. E. Gavins, Reinhold Munker. Potent inhibition of the cell proliferation and induction of apoptosis in lymphoma cells by the anthelminthic drug niclosamide: in vitro data. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4816.