NHE-1 is the sodium–hydrogen exchanger isoform present in erythroid cells

AbstractErythrocyte sodium hydrogen exchanger (NHE) represents one of a limited number of sodium entry pathway in erythrocytes. At least five NHE isoforms have been identified, differing in tissue specificity, regulatory characteristics, and pharmacological sensitivities. Although physiological characteristics of erythrocyte NHE suggest that the widely expressed NHE-1 isoform may be present, evidence is not conclusive and does not exclude the existence of other isoforms. In this study, Northern blot and reverse transcription–polymerase chain reaction (RT–PCR) analyses were used to test for five NHE isoforms in erythroid cells. Blood from patients with sickle cell disease was depleted of white blood cells (WBC) by passage through leukocyte filters and cellulose column. RT–PCR performed on WBC depleted reticulocyte RNA using a NHE-1 primer set yielded product a of expected size, the sequence of which was identical to the published human NHE-1 sequence. Northern blot analysis of the reticulocyte RNA using a 1.6 kb probe revealed a message of approximately 5.0 kb in size. RT–PCR analysis of rat kidney RNA using primers specific for NHE isoforms -2, -3, -4 and rat brain RNA using primer specific for NHE-5 isoform yielded products of expected size, whereas WBC depleted RNA under identical conditions yielded no products. These results identify the erythroid isoform of the sodium–hydrogen exchanger as NHE-1

Abstract 990: Metabolic shift in cancer by re-programming of mitochondrial machinery: Novel insight into Warburg-dependent mechanism

Abstract Otto Warburg described his seminal observation that cancer cells negotiated a switch towards anaerobic respiration from oxidative phosphorylation in return for sustained growth and evasion of normal cellular processes such as cell cycle control and apoptosis. The underlying mechanism that governs this phenomenon has yet to be firmly established. Given the centrality of mitochondria to apoptosis and bioenergetics, we proposed experiments that employed ubidecarenone as an intracellular energy ambassador wherein high-throughput genomic, metabolomic, and proteomic analyses were employed to capture the energetic and molecular signature of cancer cells. Analyses were performed on oncogenic breast, prostate, liver, pancreatic, skin (melanoma, squamous cell carcinoma) and were compared to normal fibroblasts, keratinocytes, hepatocytes, kidney, adipocytes, and human aortic and endothelial cells. The results suggested a hallmark discovery that clearly delineated a differential effect of cancer vs. normal cells under hyperglycemic, hypoxic, and lactate-stressed conditions. A decreased activation of the Pentose Phosphate Shunt was significantly downregulated in cancer cells suggestive of a shift towards utilization of oxygen and glycolysis while Bcl-2 dependent cell death mechanisms were restored. Most notably, cross-talk mechanisms related to p53 and Vegf in the cancer cells and tumor markers and pathology in animal models of melanoma and pancreatic cancer demonstrated efficacy in a topical and intravenous form of Cytotech Labs API 31510, a ubidecarenone based technology that targets the Bcl-2 protein family downstream of a cellular metabolic shift. Conversely, normal cells did not exhibit the aforementioned characteristic indicative of a cancer-specific mechanism. We propose a novel underlying mechanism for the Warburg Hypothesis and demonstrate that pre-clinical work on API 31510 in cancer suggest that it may be a viable agent for control of cancer metabolism and serve as safe, effective anti-cancer agent. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 990. doi:10.1158/1538-7445.AM2011-990</jats:p

Abstract 4321: Effect of pretreatment, dose and route of administration of BPM31510 (Coenyzme Q10 containing proprietary formulation) alone or in combination with gemcitabine improves survival in pancreatic cancer

Abstract In-vitro studies have demonstrated the ability of BPM31510, a Coenzyme Q10 (CoQ10) containing proprietary formulation to effectuate a metabolic switch from glycolysis to mitochondrial OXPHOS resulting in the activation of apoptotic pathways specifically in cancer cells. The effectiveness of BPM31510 in combination with standard of care chemotherapy agents in in-vitro and in-vivo models has been previously established. The current study was to determine the effect of dose, route of administration and combination with gemcitabine on survival in an animal model of pancreatic cancer. BPM31510 administered in three divided doses (50mg/kg vs 75mg/kg body weight daily, intravenous) was associated with dose dependent increase in survival and was significantly better when combined with gemcitabine. Continuous infusion of BPM31510 significantly improved survival rates compared to three divided doses (50mg/kg or 75mg/kg body weight) of BPM31510, with best outcomes at 200mg/kg body weight dose. Sixty day pretreatment with BPM31510 alone followed by combination with gemcitabine was also associated with improved survival outcomes with either gemcitabine or BPM31510 alone. The data suggests that dose and route of administration of BPM31510 alone or in combination with standard of care chemotherapy agents influences and improves survival in animal model of pancreatic cancer. Citation Format: Niven Narain, Lucia Mauro, Assuan Lens, Viatcheslav Akmaev, Rangaprasad Sarangarajan, Joaquin Jimenez. Effect of pretreatment, dose and route of administration of BPM31510 (Coenyzme Q10 containing proprietary formulation) alone or in combination with gemcitabine improves survival in pancreatic cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4321. doi:10.1158/1538-7445.AM2014-4321</jats:p

Current Status of Metabolomic Biomarker Discovery: Impact of Study Design and Demographic Characteristics

Widespread application of omic technologies is evolving our understanding of population health and holds promise in providing precise guidance for selection of therapeutic interventions based on patient biology. The opportunity to use hundreds of analytes for diagnostic assessment of human health compared to the current use of 10&ndash;20 analytes will provide greater accuracy in deconstructing the complexity of human biology in disease states. Conventional biochemical measurements like cholesterol, creatinine, and urea nitrogen are currently used to assess health status; however, metabolomics captures a comprehensive set of analytes characterizing the human phenotype and its complex metabolic processes in real-time. Unlike conventional clinical analytes, metabolomic profiles are dramatically influenced by demographic and environmental factors that affect the range of normal values and increase the risk of false biomarker discovery. This review addresses the challenges and opportunities created by the evolving field of clinical metabolomics and highlights features of study design and bioinformatics necessary to maximize the utility of metabolomics data across demographic groups

Parenteral CoQ10 Formulation (BPM31510) Significantly Improves Survival In Animal Model Of Leukemia Including The Resolution Of Paraplegia Due To Brain Metastasis

Abstract Leukemia cells exhibit alterations in intermediary metabolism similar to other cancers, wherein ATP sourcing is shunted from mitochondrial oxphos towards glycolytic preponderance (Warburg Effect) to meet oncogenic proliferative demands. A consequence of this metabolic switch is the simultaneous short-circuit of the programmed death pathways, leading to a immortalization program in cancer cells including leukemia. Delivery of high levels of CoQ10 in a lipid nanodispersion mixture (BPM31510) has been demonstrated to preferentially shift metabolic networks from glycolysis towards mitochondrial-centric oxphos and recapitulation of apoptotic pathways in various cancers in vitro and in vivo models. Given the centrality of the Bcl-2 involvement in the etiology of leukemia, this study focused on investigation of the effectiveness of BPM31510 in animal models of erythroid and myeloid leukemia. Human acute erythro-leukemia (K562) and acute myeloid leukemia (KG1) models were developed in immune-compromised mice. The mice (total n=120 for each model respectively) were randomized into four (n=30/group) treatment groups: Untreated (control); BPM31510 (75 mg/kg, once/day); chemotherapy (Adriamycin [5mg/kg; once/wk]+AraC [25mg/kg; 5 days] and BPM31510+chemotherapy. All dosing were intravenous and followed a protocol of 3wk treatment followed by 1wk rest. In both leukemia models, combination of BPM31510 with chemotherapy was associated with significant increase in survival compared to other groups. BPM31510 alone improved survival compared to chemotherapy in myeloid leukemia, not in erythroleukemia model. In a separate study, a rat (Fisher 344) chloroleukemia (MIA C51) model of CNS leukemia was developed that demonstrated paraplegia and urinary retention as a result of brain metastasis. Administration of BPM31510 (50 mg/kg/day, IP) was associated with complete resolution of limb paralysis demonstrating the ability of BPM31510 in penetrating into the CNS. Moreover, BPM31510 (50 mg/kg/day, IP) administration was associated with significant increase in survival in animals with metastasis to the lungs and liver. The data provides encouraging evidence of the potential translational use of CoQ10 containing BPM31510 in the treatment of leukemia. A Phase 1 study in relapse acute leukemia is to be started shortly. Disclosures: Narain: Berg: Employment. Akmaev:Berg: Employment. Benaim:Berg: Employment. Sarangarajan:Berg: Employment. Jimenez:Berg: Membership on an entity’s Board of Directors or advisory committees