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Article ; Online: Metabolic targeting of malignant tumors: small-molecule inhibitors of bioenergetic flux.

Mathupala, Saroj P

Recent patents on anti-cancer drug discovery

2010 Volume 6, Issue 1, Page(s) 6–14

Abstract: Metabolism in tumors deviates significantly from that of normal tissues. Increasingly, the underlying aberrant metabolic pathways are being considered as novel targets for cancer therapy. Denoted "metabolic targeting", small molecule drugs are under ... ...

Abstract	Metabolism in tumors deviates significantly from that of normal tissues. Increasingly, the underlying aberrant metabolic pathways are being considered as novel targets for cancer therapy. Denoted "metabolic targeting", small molecule drugs are under investigation for focused inhibition of key metabolic steps that are utilized by tumors, since such inhibitors should harbor minimal toxicity towards surrounding normal tissues. This review will examine the primary biochemical pathways that tumors harness to enhance their bioenergetic capacity, which in turn, help their rapid proliferation and metastasis within the host. It is hoped that "metabolite-mimetic" drugs can be utilized to interfere with metabolic flux pathways active within the tumor, and across tumor-microenvironment boundary. In fact, the major pathways of mammalian metabolism, i.e., the carbohydrate, amino-acid, and fatty-acid metabolic pathways have been examined as putative targets for drug development, with some drug candidates advancing to phase II/III stages. In this regard, glucose metabolism, i.e., the glycolytic pathway - that predominates the bio-energetic flux in tumors, and the associated mitochondrial metabolism have received the most attention as suitable "druggable" targets, focused either at the pathway enzymes or at the plasma-membrane-bound metabolite transporters. Outlined in this review are pre-clinical studies that have led to the discovery of promising drug candidates to target tumor-metabolic flux, and ensuing patents, with descriptions of the biochemical rationale for the combinatorial strategy of a particular metabolic pathway-drug candidate pair.
MeSH term(s)	Animals ; Antineoplastic Agents/administration & dosage ; Antineoplastic Agents/pharmacology ; Antineoplastic Agents/therapeutic use ; Energy Metabolism/drug effects ; Energy Metabolism/genetics ; Enzyme Inhibitors/therapeutic use ; Glycolysis/drug effects ; Glycolysis/genetics ; Humans ; Metabolic Networks and Pathways/drug effects ; Metabolic Networks and Pathways/genetics ; Models, Biological ; Molecular Targeted Therapy/methods ; Neoplasms/drug therapy ; Neoplasms/genetics ; Neoplasms/metabolism ; Small Molecule Libraries
Chemical Substances	Antineoplastic Agents ; Enzyme Inhibitors ; Small Molecule Libraries
Language	English
Publishing date	2010-11-29
Publishing country	United Arab Emirates
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	2250820-X
ISSN	2212-3970 ; 1574-8928
ISSN (online)	2212-3970
ISSN	1574-8928
DOI	10.2174/157489211793980114
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Article ; Online: Delivery of small-interfering RNA (siRNA) to the brain.

Mathupala, Saroj P

Expert opinion on therapeutic patents

2009 Volume 19, Issue 2, Page(s) 137–140

Abstract: Background: Two fundamental difficulties in the delivery of drugs to treat central nervous system (CNS) diseases are the systemic delivery of therapeutics across the bloodbrain-barrier (BBB), and the targeting of drugs to specific tissues or cells ... ...

Abstract	Background: Two fundamental difficulties in the delivery of drugs to treat central nervous system (CNS) diseases are the systemic delivery of therapeutics across the bloodbrain-barrier (BBB), and the targeting of drugs to specific tissues or cells within the brain. With the advent and promise of RNA-based therapeutics that utilize RNA interference (RNAi) to trigger specific silencing of genes within diseased tissues, the necessity to surmount such obstacles has become even more urgent. Objective: Most pre-clinical and clinical studies on delivery of RNAi to the CNS have utilized invasive, intra-cerebral delivery of RNA to the targeted tissue. Thus, methods need to be developed to facilitate delivery of therapeutically significant quantities of RNA to the CNS via the systemic route, and to elicit clinically significant RNAi effects within the CNS tissues. Methods: Cell-penetrating-peptides (CPPs) are 'molecular delivery vehicles' that can traverse cell membranes and co-transport peptides or polynucleotides. The present invention examines 1) the utility of CPP-RNA duplexes for delivery of RNA to CNS tissues and, 2) cell-mediated release of the RNA payload once the CPP-RNA duplex is internalized by the CNS cells. Conclusions: The invention and embodiments listed therein outline molecular tools that can be adapted for non-invasive, systemic delivery of therapeutic RNA to the CNS in a future clinical setting.
MeSH term(s)	Animals ; Blood-Brain Barrier/metabolism ; Brain/metabolism ; Drug Delivery Systems ; Drug Evaluation, Preclinical ; Gene Silencing ; Humans ; Patents as Topic ; Peptides/administration & dosage ; Peptides/pharmacokinetics ; RNA, Small Interfering/administration & dosage ; RNA, Small Interfering/pharmacokinetics
Chemical Substances	Peptides ; RNA, Small Interfering
Language	English
Publishing date	2009-05-08
Publishing country	England
Document type	Journal Article ; Review
ZDB-ID	1186201-4
ISSN	1744-7674 ; 0962-2594 ; 1354-3776
ISSN (online)	1744-7674
ISSN	0962-2594 ; 1354-3776
DOI	10.1517/13543770802680195
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Article ; Online: A Lab Assembled Microcontroller-Based Sensor Module for Continuous Oxygen Measurement in Portable Hypoxia Chambers.

Mathupala, Saroj P / Kiousis, Sam / Szerlip, Nicholas J

PloS one

2016 Volume 11, Issue 2, Page(s) e0148923

Abstract: Background: Hypoxia-based cell culture experiments are routine and essential components of in vitro cancer research. Most laboratories use low-cost portable modular chambers to achieve hypoxic conditions for cell cultures, where the sealed chambers are ... ...

Abstract	Background: Hypoxia-based cell culture experiments are routine and essential components of in vitro cancer research. Most laboratories use low-cost portable modular chambers to achieve hypoxic conditions for cell cultures, where the sealed chambers are purged with a gas mixture of preset O2 concentration. Studies are conducted under the assumption that hypoxia remains unaltered throughout the 48 to 72 hour duration of such experiments. Since these chambers lack any sensor or detection system to monitor gas-phase O2, the cell-based data tend to be non-uniform due to the ad hoc nature of the experimental setup. Methodology: With the availability of low-cost open-source microcontroller-based electronic project kits, it is now possible for researchers to program these with easy-to-use software, link them to sensors, and place them in basic scientific apparatus to monitor and record experimental parameters. We report here the design and construction of a small-footprint kit for continuous measurement and recording of O2 concentration in modular hypoxia chambers. The low-cost assembly (US$135) consists of an Arduino-based microcontroller, data-logging freeware, and a factory pre-calibrated miniature O2 sensor. A small, intuitive software program was written by the authors to control the data input and output. The basic nature of the kit will enable any student in biology with minimal experience in hobby-electronics to assemble the system and edit the program parameters to suit individual experimental conditions. Results/conclusions: We show the kit's utility and stability of data output via a series of hypoxia experiments. The studies also demonstrated the critical need to monitor and adjust gas-phase O2 concentration during hypoxia-based experiments to prevent experimental errors or failure due to partial loss of hypoxia. Thus, incorporating the sensor-microcontroller module to a portable hypoxia chamber provides a researcher a capability that was previously available only to labs with access to sophisticated (and expensive) cell culture incubators.
MeSH term(s)	Atmosphere Exposure Chambers/economics ; Brain Neoplasms/pathology ; Cell Hypoxia ; Cell Line, Tumor ; Electronics ; Equipment Design ; Fluorometry/economics ; Fluorometry/instrumentation ; Humans ; Manometry/economics ; Manometry/instrumentation ; Microcomputers/economics ; Oxygen/analysis ; Software ; Thermometry/economics ; Thermometry/instrumentation ; Tissue Culture Techniques/economics ; Tissue Culture Techniques/instrumentation
Chemical Substances	Oxygen (S88TT14065)
Language	English
Publishing date	2016
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural
ISSN	1932-6203
ISSN (online)	1932-6203
DOI	10.1371/journal.pone.0148923
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Article ; Online: A Lab Assembled Microcontroller-Based Sensor Module for Continuous Oxygen Measurement in Portable Hypoxia Chambers.

Saroj P Mathupala / Sam Kiousis / Nicholas J Szerlip

PLoS ONE, Vol 11, Iss 2, p e

2016 Volume 0148923

Abstract: Hypoxia-based cell culture experiments are routine and essential components of in vitro cancer research. Most laboratories use low-cost portable modular chambers to achieve hypoxic conditions for cell cultures, where the sealed chambers are purged with a ...

Abstract	Hypoxia-based cell culture experiments are routine and essential components of in vitro cancer research. Most laboratories use low-cost portable modular chambers to achieve hypoxic conditions for cell cultures, where the sealed chambers are purged with a gas mixture of preset O2 concentration. Studies are conducted under the assumption that hypoxia remains unaltered throughout the 48 to 72 hour duration of such experiments. Since these chambers lack any sensor or detection system to monitor gas-phase O2, the cell-based data tend to be non-uniform due to the ad hoc nature of the experimental setup.With the availability of low-cost open-source microcontroller-based electronic project kits, it is now possible for researchers to program these with easy-to-use software, link them to sensors, and place them in basic scientific apparatus to monitor and record experimental parameters. We report here the design and construction of a small-footprint kit for continuous measurement and recording of O2 concentration in modular hypoxia chambers. The low-cost assembly (US$135) consists of an Arduino-based microcontroller, data-logging freeware, and a factory pre-calibrated miniature O2 sensor. A small, intuitive software program was written by the authors to control the data input and output. The basic nature of the kit will enable any student in biology with minimal experience in hobby-electronics to assemble the system and edit the program parameters to suit individual experimental conditions.We show the kit's utility and stability of data output via a series of hypoxia experiments. The studies also demonstrated the critical need to monitor and adjust gas-phase O2 concentration during hypoxia-based experiments to prevent experimental errors or failure due to partial loss of hypoxia. Thus, incorporating the sensor-microcontroller module to a portable hypoxia chamber provides a researcher a capability that was previously available only to labs with access to sophisticated (and expensive) cell culture incubators.
Keywords	Medicine ; R ; Science ; Q
Subject code	600
Language	English
Publishing date	2016-01-01T00:00:00Z
Publisher	Public Library of Science (PLoS)
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Voltage dependent anion channel-1 (VDAC-1) as an anti-cancer target.

Mathupala, Saroj P / Pedersen, Peter L

Cancer biology & therapy

2010 Volume 9, Issue 12, Page(s) 1053–1056

MeSH term(s)	Animals ; Apoptosis ; Humans ; Mice ; Mitochondrial Membranes ; Neoplasms/drug therapy ; Neoplasms/metabolism ; RNA Interference ; RNA, Small Interfering ; Voltage-Dependent Anion Channel 1/genetics ; Voltage-Dependent Anion Channel 1/metabolism
Chemical Substances	RNA, Small Interfering ; Voltage-Dependent Anion Channel 1 (EC 1.6.-)
Language	English
Publishing date	2010-06-21
Publishing country	United States
Document type	Comment ; Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	2146305-0
ISSN	1555-8576 ; 1538-4047
ISSN (online)	1555-8576
ISSN	1538-4047
DOI	10.4161/cbt.9.12.12451
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Article ; Online: Low-cost media formulation for culture of brain tumor spheroids (neurospheres).

Monterey, Michael D / Szerlip, Nicholas J / Mathupala, Saroj P

BioTechniques

2013 Volume 55, Issue 2, Page(s) 83–88

Abstract: Recent studies have found that the biological features of primary tumors are faithfully recapitulated when a patient's tumor is processed and then maintained as a 3-D spheroid in specialized cell culture media. However, a major drawback for maintenance ... ...

Abstract	Recent studies have found that the biological features of primary tumors are faithfully recapitulated when a patient's tumor is processed and then maintained as a 3-D spheroid in specialized cell culture media. However, a major drawback for maintenance and routine passage of primary tumors as spheroids has been the high cost of custom-formulated media compared to regular serum-supplemented media. Here we report the formulation of a cost-effective, serum-free medium in which high-grade primary brain tumor (glioblastoma) explants can be established and maintained as spheroids. Based on DMEM, this formulation requires only supplementation with several amino acids, vitamins, synthetic EGF, and bFGF, with most of the cost being associated with the growth factors. A simple addition of BSA (fraction V) obviated the need for numerous other components (or human serum) commonly used in the specialized commercial media formulations.
MeSH term(s)	Brain Neoplasms/pathology ; Cell Culture Techniques ; Culture Media, Serum-Free/economics ; Glioblastoma/pathology ; Humans ; Phenotype ; Reproducibility of Results ; Spheroids, Cellular/cytology ; Tumor Cells, Cultured/cytology
Chemical Substances	Culture Media, Serum-Free
Language	English
Publishing date	2013-08-12
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	48453-2
ISSN	1940-9818 ; 0736-6205
ISSN (online)	1940-9818
ISSN	0736-6205
DOI	10.2144/000114066
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Article: The pivotal roles of mitochondria in cancer: Warburg and beyond and encouraging prospects for effective therapies.

Mathupala, Saroj P / Ko, Young H / Pedersen, Peter L

Biochimica et biophysica acta

2010 Volume 1797, Issue 6-7, Page(s) 1225–1230

Abstract: Tumors usurp established metabolic steps used by normal tissues for glucose utilization and ATP production that rely heavily on mitochondria and employ a route that, although involving mitochondria, includes a much greater dependency on glycolysis. First ...

Abstract	Tumors usurp established metabolic steps used by normal tissues for glucose utilization and ATP production that rely heavily on mitochondria and employ a route that, although involving mitochondria, includes a much greater dependency on glycolysis. First described by Otto Warburg almost nine decades ago [1], this aberrant phenotype becomes more pronounced with increased tumor malignancy [2]. Thus, while maintaining their capacity for respiration, tumors "turn more parasitic" by enhancing their ability to scavenge glucose from their surroundings. With excess glucose at hand, tumors shunt their metabolic flux more toward glycolysis than do their normal cells of origin, a strategy that allows for their survival when oxygen is limiting while providing them a mechanism to poison their extra-cellular environment with acid, thus paving the way for invasion and metastasis. Significantly, tumors harness a crucial enzyme to regulate and support this destructive path--to entrap and channel glucose toward glycolysis. This enzyme is an isoform of hexokinase, referred to as hexokinase type II, and also in abbreviated form as HK-2 or HK II. Due to many-faceted molecular features at genetic, epigenetic, transcriptional, and enzymatic levels, including sub-cellular localization to mitochondria, HK-2 facilitates and promotes the high glycolytic tumor phenotype [3]. Thus, HK-2 represents a pivotal model gene or enzyme that tumors "select for" during tumorigenesis in order to facilitate their destructive path. In this review, we examine the roles played by mitochondrial bound HK-2 within the context of the highly choreographed metabolic roulette of malignant tumors. Recent studies that outline how the aberrant glycolytic flux can be subverted toward a more "normal" metabolic phenotype, and how the glycolytic flux affects the tumor microenvironment to facilitate tumor dissemination are also described, including how these very features can be harnessed in new metabolic targeting strategies to selectively debilitate tumors.
MeSH term(s)	Animals ; Glucose/metabolism ; Glycolysis ; Hexokinase/genetics ; Hexokinase/metabolism ; History, 20th Century ; History, 21st Century ; Humans ; Lactic Acid ; Mitochondria/metabolism ; Models, Biological ; Monocarboxylic Acid Transporters/metabolism ; Neoplasms/genetics ; Neoplasms/metabolism ; Neoplasms/pathology ; Neoplasms/therapy ; Proton Pumps/metabolism ; Research/history
Chemical Substances	Monocarboxylic Acid Transporters ; Proton Pumps ; Lactic Acid (33X04XA5AT) ; Hexokinase (EC 2.7.1.1) ; Glucose (IY9XDZ35W2)
Language	English
Publishing date	2010-04-08
Publishing country	Netherlands
Document type	Historical Article ; Journal Article ; Review
ZDB-ID	60-7
ISSN	1879-2596 ; 1879-260X ; 1872-8006 ; 1879-2642 ; 1879-2618 ; 1879-2650 ; 0006-3002 ; 0005-2728 ; 0005-2736 ; 0304-4165 ; 0167-4838 ; 1388-1981 ; 0167-4889 ; 0167-4781 ; 0304-419X ; 1570-9639 ; 0925-4439 ; 1874-9399
ISSN (online)	1879-2596 ; 1879-260X ; 1872-8006 ; 1879-2642 ; 1879-2618 ; 1879-2650
ISSN	0006-3002 ; 0005-2728 ; 0005-2736 ; 0304-4165 ; 0167-4838 ; 1388-1981 ; 0167-4889 ; 0167-4781 ; 0304-419X ; 1570-9639 ; 0925-4439 ; 1874-9399
DOI	10.1016/j.bbabio.2010.03.025
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Article ; Online: Low-cost media formulation for culture of brain tumor spheroids (neurospheres)

Michael D. Monterey / Nicholas J. Szerlip / Saroj P. Mathupala

BioTechniques, Vol 55, Iss 2, Pp 83-

2013 Volume 88

Abstract	Recent studies have found that the biological features of primary tumors are faithfully recapitulated when a patient's tumor is processed and then maintained as a 3-D spheroid in specialized cell culture media. However, a major drawback for maintenance and routine passage of primary tumors as spheroids has been the high cost of custom-formulated media compared to regular serum-supplemented media. Here we report the formulation of a cost-effective, serum-free medium in which high-grade primary brain tumor (glioblastoma) explants can be established and maintained as spheroids. Based on DMEM, this formulation requires only supplementation with several amino acids, vitamins, synthetic EGF, and bFGF, with most of the cost being associated with the growth factors. A simple addition of BSA (fraction V) obviated the need for numerous other components (or human serum) commonly used in the specialized commercial media formulations.
Keywords	Neurosphere ; glioma ; spheroid ; CD133 ; serum-free culture ; Biology (General) ; QH301-705.5
Language	English
Publishing date	2013-08-01T00:00:00Z
Publisher	Future Science Ltd
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Hexokinase-2 bound to mitochondria: cancer's stygian link to the "Warburg Effect" and a pivotal target for effective therapy.

Mathupala, Saroj P / Ko, Young H / Pedersen, Peter L

Seminars in cancer biology

2008 Volume 19, Issue 1, Page(s) 17–24

Abstract: ... nutrient inside the tumor cells as glucose-6-P, some of which is funneled off to serve as carbon precursors ...

Abstract	The most common metabolic hallmark of malignant tumors, i.e., the "Warburg effect" is their propensity to metabolize glucose to lactic acid at a high rate even in the presence of oxygen. The pivotal player in this frequent cancer phenotype is mitochondrial-bound hexokinase [Bustamante E, Pedersen PL. High aerobic glycolysis of rat hepatoma cells in culture: role of mitochondrial hexokinase. Proc Natl Acad Sci USA 1977;74(9):3735-9; Bustamante E, Morris HP, Pedersen PL. Energy metabolism of tumor cells. Requirement for a form of hexokinase with a propensity for mitochondrial binding. J Biol Chem 1981;256(16):8699-704]. Now, in clinics worldwide this prominent phenotype forms the basis of one of the most common detection systems for cancer, i.e., positron emission tomography (PET). Significantly, HK-2 is the major bound hexokinase isoform expressed in cancers that exhibit a "Warburg effect". This includes most cancers that metastasize and kill their human host. By stationing itself on the outer mitochondrial membrane, HK-2 also helps immortalize cancer cells, escapes product inhibition and gains preferential access to newly synthesized ATP for phosphorylating glucose. The latter event traps this essential nutrient inside the tumor cells as glucose-6-P, some of which is funneled off to serve as carbon precursors to help promote the production of new cancer cells while much is converted to lactic acid that exits the cells. The resultant acidity likely wards off an immune response while preparing surrounding tissues for invasion. With the re-emergence and acceptance of both the "Warburg effect" as a prominent phenotype of most clinical cancers, and "metabolic targeting" as a rational therapeutic strategy, a number of laboratories are focusing on metabolite entry or exit steps. One remarkable success story [Ko YH, Smith BL, Wang Y, Pomper MG, Rini DA, Torbenson MS, et al. Advanced cancers: eradication in all cases using 3-bromopyruvate therapy to deplete ATP. Biochem Biophys Res Commun 2004;324(1):269-75] is the use of the small molecule 3-bromopyruvate (3-BP) that selectively enters and destroys the cells of large tumors in animals by targeting both HK-2 and the mitochondrial ATP synthasome. This leads to very rapid ATP depletion and tumor destruction without harm to the animals. This review focuses on the multiple roles played by HK-2 in cancer and its potential as a metabolic target for complete cancer destruction.
MeSH term(s)	Adenosine Triphosphate/metabolism ; Animals ; Energy Metabolism/physiology ; Gene Expression Regulation, Neoplastic/genetics ; Glucose/metabolism ; Glycolysis/physiology ; Hexokinase/metabolism ; Humans ; Mitochondria/enzymology ; Mitochondria/metabolism ; Neoplasms/drug therapy ; Neoplasms/enzymology ; Neoplasms/genetics ; Neoplasms/metabolism ; Phosphorylation/physiology ; Protein Isoforms/metabolism ; Pyruvates/metabolism ; Pyruvates/therapeutic use
Chemical Substances	Protein Isoforms ; Pyruvates ; bromopyruvate (63JMV04GRK) ; Adenosine Triphosphate (8L70Q75FXE) ; Hexokinase (EC 2.7.1.1) ; Glucose (IY9XDZ35W2)
Language	English
Publishing date	2008-12-03
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	1033980-2
ISSN	1096-3650 ; 1044-579X
ISSN (online)	1096-3650
ISSN	1044-579X
DOI	10.1016/j.semcancer.2008.11.006
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Article ; Online: An agarose-based cloning-ring anchoring method for isolation of viable cell clones

Saroj P. Mathupala / Andrew E. Sloan

BioTechniques, Vol 46, Iss 4, Pp 305-

2009 Volume 307

Abstract: Isolation of clonal cell populations is a crucial aspect of cell biology during engineering of specific cell strains with both genotypic and phenotypic variations. The use of cloning rings is the most established method, but requires anchoring chemicals ... ...

Abstract	Isolation of clonal cell populations is a crucial aspect of cell biology during engineering of specific cell strains with both genotypic and phenotypic variations. The use of cloning rings is the most established method, but requires anchoring chemicals or material that can often interfere with quantitative clonal-cell isolation and causes physical damage to the cells. Here we report a non-toxic, cell culture–compatible method that uses aga-rose for embedding the cloning rings during isolation of cell clones from monolayer cultures, with enhanced cell-viability and reproducibility during downstream applications. The method is simple and rapid, minimizing the chances for desiccation or cross-contamination during colony-lifts.
Keywords	cloning-rings ; cell cloning ; tissue culture ; clonal isolation ; mammalian cells ; cell culture ; Biology (General) ; QH301-705.5
Language	English
Publishing date	2009-04-01T00:00:00Z
Publisher	Future Science Ltd
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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