New Therapy for Brain Tumors: High-Pressure Oxygen Rewires Glucose Metabolism in Glioblastomas

Glioblastoma (GBM) is an aggressive type of brain tumor, and one of the deadliest cancers. GBM is often treated with surgery, radiation and chemotherapy, but even if the initial treatment is successful, a majority of patients relapse within months. One reason why GBM is so difficult to treat is the hypoxic (low-oxygen) tumor environment. It is known that hypoxic cells are resistant to radiotherapy; the greater the number of tumor stem cells in a hypoxic environment, the less efficient radiotherapy is at controlling tumor growth.

A new therapeutic approach aims to remove the hypoxic environment in GBM by administering pure oxygen to patients at high pressure, known as “hyperbaric oxygen (HBO) therapy”. Previous studies have shown that HBO improves the efficacy of radiotherapy in GBM patients. However, the therapeutic mechanism of HBO was largely unknown. That is, until now.

Dr. Anna Tesei, the Head of Radiobiomics and Drug Discovery at the Biosciences Laboratory of IRST-IRCCS in Italy, recently published a study on the mechanism in which HBOT affects GBM tumor cells and the tumor environment. “The main purpose of our study was to provide a preclinical rationale for the use of hyperbaric oxygen in association with radiotherapy for the treatment of GBM,” she says.

Dr. Anna Tesei is studing glucose metabolism in glioblastoma cells.
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LncRNA: The Long and Short of “Junk RNA”

The Central Dogma and Junk DNA

lncRNA, long noncoding RNA

On September 19, 1957, Francis Crick delivered a lecture during a symposium at University College London, titled “Protein Synthesis”. The lecture was published a year later (1); in it, Crick quotes his colleague James Watson as saying, “The most significant thing about the nucleic acids is that we don’t know what they can do.” In contrast, Crick argued that proteins play a central, indispensable role as enzymes within the cell that catalyze a variety of chemical reactions. He believed that the main role of genetic material was to control the synthesis of proteins, although the mechanism of that process was not known.

Crick’s hypothesis came to be known as the central dogma of molecular biology, and it was immortalized in his hand-written notes that described the flow of information from DNA to RNA to proteins. This achievement was all the more remarkable, considering that messenger RNAs were completely unknown at that time, and very little was known about how the cellular translational machinery functioned within the cytoplasm to synthesize proteins. Although the later discovery of retroviruses appeared to challenge Crick’s central dogma, he explained quite succinctly that his original statement had simply been misunderstood, and that information could flow in both directions between DNA and RNA (2).

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The NLRP3 Inflammasome: Flipping the Switch

It’s been just over 10 years since the world lost a pioneering immunologist and biochemist, Dr. Jürg Tschopp. He died tragically during a hiking trip in the Swiss Alps on March 22, 2011. A host of academic journals, including Science, Nature and Cell, paid tribute to Dr. Tschopp with obituaries that highlighted his many accomplishments in the fields of apoptosis and immunology.

In 2002, a team led by Dr. Tschopp at the University of Lausanne, Switzerland, was studying the role of the proinflammatory cytokine interleukin 1 beta (IL-1β). This cytokine is produced in the cytoplasm as an inactive precursor (pro-IL-1β). It is cleaved by caspase-1 to the active form, but the exact process by which caspase-1 itself is activated was unknown at the time. Several members of the caspase family contain a conserved region known as the caspase recruitment domain or CARD, and it was proposed that this domain was essential to caspase activation.

Based on similarity to another protein containing an N-terminal CARD motif (Apaf-1) that is involved in activation of caspase-9, the researchers examined the roles of a family of proteins known as NALP1, NALP2 and NALP3 (1). In particular, they were interested in NALP1, which is involved in the immune response. Unlike Apaf-1, NALP1 contains a CARD motif at the C terminus, while the N terminus contains a related motif known as a pyrin-like domain (PYD). The research team had previously showed that the PYD region of NALP1 interacted with an adapter protein known as PYCARD or ASC, which also contains an N-terminal PYD and C-terminal CARD.

The results of the team’s in vitro binding, activation and immunodetection studies showed that a multi-unit protein complex is responsible for caspase activation, and they called this complex the “inflammasome” (1). It is composed of caspase-1, caspase-5, PYCARD/ASC and NALP1.

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RAS-Targeted Drug Discovery: From Challenge to Opportunity

cancer cell, ras-targeted drug discovery

In 1963, Jennifer Harvey was studying Moloney murine leukemia virus (MMLV) at the cancer research department of the London Hospital Research Laboratories. After routine transfers of plasma from MMLV-infected rats to mice, she made an unusual discovery. In addition to the expected leukemia, the mice that received the plasma developed solid tumors (soft-tissue sarcomas), primarily in the spleen (1). A few years later, Werner Kirsten at the University of Chicago observed similar results working with mouse erythroblastosis virus (MEV) (2).

Subsequent research, with the advent of genome sequencing, showed that a cellular rat gene had been incorporated into the viral genome in both cases (3). These genomic sequences contained a mutation later shown to be responsible for the development of sarcomas, and the word “oncogene” became a common part of the vocabulary in cancer publications during the early 1980s (4). Harvey’s discovery led to the naming of the corresponding rat sarcoma oncogene as HRAS, while Kirsten’s related oncogene was named KRAS. Several laboratories, working independently, cloned the human homolog of the viral HRAS gene in 1982 (3). The human KRAS gene was cloned shortly thereafter, as well as a third RAS gene, named NRAS (3). Additional studies showed that a single point mutation in each of these genes led to oncogenic activation, and they have been popular targets for anticancer drug discovery efforts ever since.

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Antibody Response Differ in Adults, Kids and Potential Cross-Reactive Coronavirus Antibodies

B cell and B cell receptor cartoon. The B cell receptors are important for antibody response.
Drawing of a B cell and the B cell receptor. The receptor shows the characteristic Y shape of an immunoglobulin molecule.

B cells are the immune cells that produce antibodies (immunoglobulins or Ig) to detect intruding pathogens. B cells produce a variety of classes of antibodies. Generally during an immune response to a pathogen, whether viral or bacterial, B cells produce immunoglobulins (Ig) IgM and IgD, and later in the response, IgG and IgA, that are specific to the intruding organism. These Igs capture and aid in neutralizing the pathogen.

Ig classes can be studied by sequencing the B cell receptor (BCR), which binds antigen specifically. BCRs are formed via irreversible gene segment rearrangements of variable, diversity and joining (VDJ) genes. Ig classes can be diversified through somatic hypermutation and class-switch recombination of these gene segments (1).

B cell receptors with high sequence similarity can be found in individuals exposed to the same antigen, demonstrating that antigen exposure can result in similar B cell clones and memory B cells between individuals, both adults and children (1).

However, B cell immune responses can differ between adults and children. For example, children use more B cell clones that form neutralizing antibodies to HIV-1. And children infected with SARS-CoV-2 generally have milder illness than infected adults. SARS-CoV-2-infected children have lower antibody titers to the virus and more IgG-specific response to SARS-CoV-2 spike protein than to the nucleocapsid protein (1). These differences can contribute to faster SARS-CoV-2 clearance and lower viral loads in children versus adults.

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NAD: A Renaissance Molecule and its Role in Cell Health

Promega NAD/NADH-Glo system and how to prepare samples for  identification of NAD or NADH.
Promega NAD/NADH-Glo system and how to prepare samples for identification of NAD or NADH.

NAD is a pyridine nucleotide. It provides the oxidation and reduction power for generation of ATP by mitochondria. For many years it was believed that the primary function of NAD/NADH in cells was to harness and transfer energy  from glucose, fatty and amino acids through pathways like glycolysis, beta-oxidation and the citric acid cycle.

Today, however, NAD is recognized as an important cell signaling molecule and substrate. The many regulatory pathways now known to use NAD+ in signaling include multiple aspects of cellular homeostasis, energy metabolism, lifespan regulation, apoptosis, DNA repair and telomere maintenance.

This resurrection of NAD importance is due in no small part to the discovery of NAD-using enzymes, especially the sirtuins. Continue reading “NAD: A Renaissance Molecule and its Role in Cell Health”

Bartonella sp. and Your Cat (or Dog, Horse, Rat): An Emerging Infectious Disease

If we’ve learned nothing else since February or March of 2020, we’ve learned that emerging infectious diseases are a real threat to human health globally. In a bad news/good news kind of way, Bartonellosis is an emerging infectious disease; however, it’s not spread by airborne droplets or respiration.

But if any of your family pets bring a flea or tick into the house, or if you live in proximity to mice, rats, ground squirrels, rabbits, sheep, horses or cattle–you could be at risk.

Bartonella sp. is a Gram negative, rod-shaped bacteria that has been around since ancient times. It’s the bacteria responsible for cat scratch disease (1) and for Trench fever (2), which affected soldiers during WWs I and II, and affects people living in over-crowded, unsanitary conditions around the world today.

Bartonella henselae bacteria, the causative agent of cat-scratch disease or bartonellosis,. 3D illustration
Bartonella henselae bacteria, the causative agent of cat-scratch disease or bartonellosis

Bartonella sp. are known to be spread by vectors such as fleas, which are part of the transmission cycle for cat scratch disease and the human body louse, the vector for transmission of Trench fever (3).

This animal-to-human transmission of Bartonella sp. classifies it as a zoonosis.

Infection due to Bartonella sp. often appear to be self-limiting, such as swelling in regional lymph nodes due to a cat scratch disease. In such cases, symptoms can subside without intervention. But Bartonella sp. have a nasty habit of hiding in red blood cells and in cells lining blood vessels, where they can remain undetected for a substantial period of time. This hiding place affects a host’s ability to mount an immune response, as well as affecting the ability of antibiotics to attack the bacteria.

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HPK1 Identified as Emerging Immuno-oncology Drug Target

Antibody-based immune checkpoint inhibitors remain a major focus of immuno-oncology drug research and development efforts because of their recent success in providing long-term anti-tumor responses. However, the range of response of different tumor types to these drugs is hugely varied. Small molecule kinase inhibitors that block signaling pathways involved in regulation of tumor immunity at multiple points in the “cancer immunity cycle” may provide alternate, effective therapeutics. One kinase that may be a target for such small molecule inhibitors is Hematopoietic Progenitor Kinase 1 or HPK1; the potential of this kinase as a therapeutic target was reviewed by Sawasdikosol and Burakoff (1). HPK1, also known as MAP4K1, is a member of the MAP kinase protein kinase family that negatively regulates signal transduction in T-cells, B-cells and dendritic cells of the immune system.

Artist rendering of what target engagement might look like for kinases like HPK1.
NanoBRET™ Target Engagement Assay (artist rendering)
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COVID-19 Therapies: Are We There Yet?

A year after COVID-19 was declared a pandemic, collaborative efforts among pharma/biotech and academic researchers have led to remarkable progress in vaccine development. These efforts include novel mRNA vaccine technology, as well as more conventional approaches using adenoviral vectors. While vaccine deployment understandably has captured the spotlight in the fight against COVID-19, there remains an urgent need to develop therapeutic agents directed against SARS-CoV-2.

COVID-19 therapeutic drugs

In the March 12 issue of Science, an editorial by Dr. Francis Collins, director of the U.S. National Institutes of Health (NIH), examines lessons learned over the past 12 months (1). Collins points out that many clinical trials of potential therapeutics were not designed to suit a public health emergency. Some were poorly designed or underpowered, yet they received considerable publicity—as was the case with hydroxychloroquine. Collins advises developing antiviral agents targeted at all major known classes of pathogens, to head off the next potential pandemic before it becomes one. A news feature in the same issue discusses the current state of coronavirus drug development (2).

The present crop of drug candidates is remarkably diverse, including repurposed drugs that were originally developed to treat diseases quite different from COVID-19. Typically, however, the mainstream candidates belong to two broad classes: small-molecule antiviral agents and large-molecule monoclonal antibodies (mAbs).

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Fighting Plant Pathogens Worldwide with the Maxwell® RSC PureFood GMO and Authentication Kit

Among the one trillion or so species that share space on our planet, complex relationships have emerged over time. Such relationships, in which two or more species closely interact, are collectively termed symbiosis. Although it’s commonly assumed that symbiotic relationships are mutually beneficial, this example constitutes only one type of symbiosis (known as mutualism). The traditional predator-prey relationship, clearly a one-sided arrangement, is also an example of symbiosis.

Olive trees in Italy are being affected by the plant pathogen Xylella fastidiosa

The sheer diversity of microbial species has led to the development of many well-characterized relationships with plants and animals. Perhaps the best-known example of mutualism in this context is the process of nitrogen fixation. In this process, various types of bacteria that live in water, soil or root nodules convert atmospheric nitrogen into forms that are readily used by plants. On the other hand, some types of bacteria-plant relationships are parasitic: the bacteria rely on the plant for survival but end up damaging their host. Parasitic relationships can have devastating ecological and economic consequences when they affect food crops.

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