Cell Biology

The Role of Bioassays in Testing New Therapeutics for Canine Cancer

Posted on by Promega
Immunoassays are bringing new hope to the treatment of canine cancer.

Every dog owner fears the day they might hear the word “cancer” from their vet. This devastating disease affects not only humans but our canine companions as well. Veterinary scientists and clinicians are now employing the same methods as researchers studying human cancer, bringing the tools of personalized cancer treatment and drug research and development to bear on canine cancer, and in the not-too-distant future the treatment for a dog’s cancer may become as personalized as the bond they share with their owner.

Developing and testing new drugs and therapies is crucial to improving cancer treatments for canines. One of the most powerful tools in the drug development toolbox is the bioassay. Bioassays enable scientists to measure the biological activity of a potential treatment compound to determine if it might be effective as a therapeutic agent. For researchers focused on advancing canine cancer therapies, bioassays are indispensable. They offer precise insights into how new drugs interact with cancer cells and the immune system.

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Neurons’ Role in FBP2 Regulation

Posted on by Kari Kenefick

Neuronal extracellular vesicles (NEVs) play a significant role in the communication between neurons and astrocytes, particularly by influencing metabolic processes such as glycolysis and lactate production. NEVs carry signaling molecules that affect the expression, degradation and oligomeric state of fructose 1,6-bisphosphatase 2 (Fbp2) in astrocytes, altering their metabolism (1).

Basic Backstory on CNS Architecture
The central nervous system (CNS) is composed of an intricate cellular communications complex, divided generally into neurons and glial cells. Neurons form the electrical signaling network, with dendrites receiving and integrating signals via chemical synapses, and axons, some up to 1 meter in length, rapidly transmitting the signals.

Glial cells, including astrocytes, microglia and other cells, interact with neuronal cells to sustain this network. For example, glial cells regulate synapse formation and provide metabolic support to promote CNS homeostasis. Glial cell dysfunction contributes to most neural diseases and can even drive neurodegenerative processes (2).

What are Neuronal Extracellular Vesicles (NEVs)?
NEVs are formed by neurons via endocytosis and are released into the extracellular space where they interact with astrocytes. These transport vesicles carry a variety of molecules, including proteins and RNA, that influence cellular processes in recipient astrocytes.

NEV and Astrocyte Interactions
Fbp2 is an important enzyme involved in glycogen synthesis that also has nonenzymatic functions, including support of neuronal processes like long-term potentiation (LTP). LTP underlies synaptic strength and plasticity and is important in both learning and memory formation.

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Visualize Protein:Protein Interactions with Bioluminescence Imaging

Posted on by Simon Moe

If you’re familiar with bioluminescence, you’ve probably used it in plate-based experiments to track various biological processes. You understand it provides distinct advantages over traditional fluorescence assays, particularly when it comes to sensitivity. However, there’s always that one nagging question: how representative is the signal on a cell-to-cell level?

Traditional approaches to decipher cell-to-cell signal rely on complex, time-intensive measures that only approximated the findings acquired through bioluminescence. That’s where the GloMax® Galaxy Bioluminescence Imager comes in. This new tool will enhance your ability to visualize proteins using NanoLuc® technology, going beyond simple numeric outputs to reveal what’s happening in individual cells.

NanoLuc® technology is well-known for its ability to assist in detecting subtle protein interactions in complex biological systems. This bright luminescent enzyme enables a much broader linear range than fluorescence, improving detection of small changes in protein activity, such as proteins interacting. Microplate readers measuring NanoLuc® assays rely on signal generated from many cells. This results in an approximation of what is occurring biologically. Truly validating those luminescent readings within a cell population has been challenging—until now. The GloMax® Galaxy allows you to perform bioluminescence imaging, moving beyond the numbers, offering the power to visualize protein interactions directly.

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Understanding the Promise of Immunotherapy in Veterinary Medicine

Posted on by Promega

Immunotherapy in veterinary medicine is a rapidly evolving field that leverages the immune system to fight diseases. These therapies are particularly effective in treating various cancers, including lymphomas, mast cell tumors, melanomas, and osteosarcomas. Beyond cancer, immunotherapies are also being explored for their potential in managing chronic inflammatory diseases, such as autoimmune disorders where the immune system mistakenly attacks the body’s own tissues. While traditionally, veterinary treatments have focused on surgery, chemotherapy, and radiation, the advent of immunotherapy offers a more targeted approach, particularly for conditions like cancer.  

This targeted approach not only minimizes collateral damage to healthy tissues but also offers the potential for longer-lasting protection by training the immune system to recognize and fight off recurrence of the disease. The interest in immunotherapies has grown in tandem with advancements in human oncology, leading to a crossover of technologies and methodologies into veterinary applications. 

How Does Immunotherapy Work?

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The Marvel of Malate: A Crucial Component in Cellular Energy Metabolism

Posted on by Simon Moe

Today’s blog written by guest author Kim Haupt.

Cellular energy metabolism is a complex biological process that relies on a suite of metabolites, each with distinct roles to maintain. Malate is one of these metabolites and is essential for maintaining cellular function through important roles in both energy production and redox homeostasis. In this blog, we highlight malate’s diverse roles and uncover some of its connections to human disease. 

Illustration of energy metablism in cell.
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Fluorescent Ligands in Biological Research: Where We’ve Been, Where We’re Headed

Posted on by Kari Kenefick

Fluorescent tags (fluorophores), have become excellent tools for labeling cells and cellular components. They can be used for imaging large molecules like proteins, on down to cellular components and enzymes such as transcription factors. Once labeled, these molecules can be tracked in tissue or inside a cell, when the right tag is used.

What is the ‘right’ tag? It’s a tag with bright signal, with low background and good photostability. For small cell components like organelles, the tag must be cell-permeable and small enough to not interfere with normal cellular processes such as transcription and metabolism.

Significant advances have been made in fluorescent tags in the past two decades. Here we look at several papers noting these advances.

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Immunometabolism: The Dynamic Interplay of Cytokines and Metabolites

Posted on by Kendra Hanslik

Immunometabolism is the study of how metabolic processes influence immune cell functions and how immune responses, in turn, shape cellular metabolism. This field examines the roles of cytokines and metabolites, which act as signaling molecules and energy sources, respectively. Cytokines can trigger or modulate metabolic pathways in immune cells, affecting their activation, growth, and response capabilities. Similarly, metabolites provide the necessary energy and building blocks that enable immune cells to proliferate, function optimally, and sustain their activity during immune responses. This dynamic interplay is crucial for maintaining health and combating disease. Together, cytokines and metabolites orchestrate a complex network that links metabolic health with immune competence on a systemic and cellular level. This blog discusses how cytokines and metabolites not only influence but also drive immune cell functions, revealing new avenues for therapeutic interventions across a range of diseases.

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Rethinking Cell Proliferation Assays

Posted on by Simon Moe

In the field of cancer research, accurately measuring cell proliferation is crucial for assessing the efficacy of therapeutic agents. This is particularly difficult with CDK 4/6 inhibitors, which arrest cells in the G1 phase without stopping their growth. This continued growth can skew results from proliferation assays which rely on factors that naturally scale with cell growth. These include mitochondrial activity (ATP levels), total cell protein, or mRNA as measured through the PRISM molecular barcoding strategy. Even though these cells are not dividing, the increase in these measurements can misleadingly suggest active proliferation.

There is growing awareness among researchers of these challenges.  A recent study highlights these limitations by demonstrating the discrepancies that arise when using metabolic assays to assess cell proliferation after treatment with drugs that induce cell cycle arrest. This blog post delves into the study’s implications and demonstrates how one of Promega’s latest developments is poised to address these challenges.

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Decoding the NAD+/NADH Ratio and Its Crucial Role in Cell Health

Posted on by Kendra Hanslik

Nicotinamide adenine dinucleotide (NAD) exists in two forms in the cell: NAD+ (oxidized) and NADH (reduced). This molecule plays a pivotal role in metabolic processes, serving as a key electron carrier in the redox reactions that drive cellular metabolism. The balance between these two forms, commonly expressed as the NAD+/NADH ratio, is crucial for maintaining cellular function and the intracellular redox state. This article explores the significance of this ratio, how it impacts cellular processes, and the consequences of NAD+/NADH ratio dysregulation.

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Cancer Moonshot: Solving Tough Problems

Posted on by Michele Arduengo

At the American Association for Cancer Research meeting in April 2016, then Vice President of the United States, Joe Biden, revealed the Cancer Moonshot℠ initiative— a program with the goals of accelerating scientific discovery in cancer research, fostering greater collaboration among researchers, and improving the sharing of data (1,2). The Cancer Moonshot is part of the 21st Century Cures Act, which earmarked $1.8 billion for cancer-related initiatives over 7 years.  The National Cancer Institute (NCI) and the Cancer Moonshot program have supported over 70 programs and consortia, and more than 250 research projects.  According to the NCI, the initiative from 2017 to 2021 resulted in over 2,000 publications, 49 clinical trials and more than 30 patent filings. Additionally, the launch of trials.cancer.gov has made information about all cancer research trials accessible to anyone who needs it (3).

“We will build a future where the word ‘cancer’ loses its power.”

First Lady, Dr. Jill Biden

In February 2022, the Biden White House announced a plan to “supercharge the Cancer Moonshot as an essential effort of the Biden-Harris administration” (4).  Biden noted in his address that, in the 25 years following the Nixon administration’s enactment of the National Cancer Act in 1971, significant strides were made in understanding cancer. It is now recognized not as a single disease, but as a collection comprising over 200 distinct diseases. This period also saw the development of new therapies and enhancements in diagnosis. However, despite a reduction in the cancer death rate by more than 25% over the past 25 years, cancer continues to be the second leading cause of death in the United States [4].

The Cancer Moonshot is a holistic attempt to improve access to information, support and patient experiences, while fostering the development of new therapeutics and research approaches to studying cancer. In this article, we will focus on research, diagnostics and drug discovery developments.

Solving for Undruggable Targets

KRAS , a member of the RAS family, has long been described as “undruggable” in large part because it is a small protein with a smooth surface that does not present many places for small molecule drugs to bind. The KRAS protein acts like an off/on switch depending upon whether it has GDP or GTP bound.  KRAS mutations are associated with many cancers including colorectal cancer (CRC), non-small cell lung cancer (NSCLC), and pancreatic ductal adenocarcinoma (PDAC). The G12 position in the protein is the most commonly mutated; G12C accounts for 13% of the mutations at this site, and is the predominant substitution found in NSCLC, while G12D is prevalent in PDAC (5).

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