National ALS Biorepository Current Research

Table of Projects

Research approved for the use of National ALS Biorepository samples.
This list will be updated as new research proposals are approved by ATSDR.

ALS Research Projects Funded by the ATSDR’s National ALS Registry

Validation of Splicing-derived ALS targets
The majority of amyotrophic lateral sclerosis (ALS) cases are sporadic (sALS) meaning the cause is unknown. Alternative splicing (AS) deregulation, which is a key driver of ALS associated with most of the major ALS genes results in aberrant AS isoforms that can be targeted with splice-switching oligonucleotides (SSO) which offers promise for the treatment of ALS. Current SSO treatments do not apply to the majority of patients with sALS. Envisagenics has developed a commercially validated SpliceCore®, an innovative software platform for the discovery of AS-based sALS targets and the development of SSOs. In this project we will validate SpliceCore identified AS targets in tissues from ALS patients.

Plasma-derived astrocyte-enriched exosomes as a platform for ALS patient stratification
The molecular mechanisms that lead to cell death in the central nervous system of ALS patients fall into three broad categories. Currently we cannot determine which mechanism is active in a given patient. Our proposal aims at developing such blood test that allows us to do identify these mechanisms. The test is based on the analysis of ultra-small vesicles that are shed by central nervous system cells and are detectable in blood. If successful, this will allow us to group ALS patients by their molecular mechanism and to conduct clinical trials with medications that are tailored to a specific mechanism. This approach can pave the way for developing effective ALS therapies and for personalized medicine.

Translating prognostic biomarkers for Amyotrophic Lateral Sclerosis
ALS is a neurodegenerative disease that progresses differently in each patient. The goal of this work is to identify specific proteins and fats in cerebrospinal fluid. This can be used to better determine how an individual will be affected by ALS. We hope to provide clinicians with easily measurable biomarkers that would help patients better understand their own disease progression.

Identification of Novel Biomarkers for ALS and characterization of The Effects of TDP-43 Mutations on Protein Function and Localization
Tissue samples from ALS patients will be analyzed for any molecular changes that occur as a result of abnormal accumulation of the protein, TDP43. We will characterize the abnormal forms of TDP43 and measure the changes in gene and protein expression that occur due to its accumulation.

Immune Dysfunctions as a Biomarker and Therapeutic Target in ALS
Amyotrophic Lateral Sclerosis (ALS) is characterized by the progressive death of motor neurons, which severely impacts the functional ability of patients in several ways, including movement of arms and legs, speech, swallowing and, eventually, breathing. There is no treatment or cure for ALS. Researchers have focused their efforts over the years on neurons, though more recent studies have shown evidence of interaction between the central nervous and immune systems, long considered separate compartments. We previously made the unexpected discovery that the adaptive immune system, which builds up the body’s protection as it is exposed to foreign pathogens, is linked to a rare form of ALS, ALS4. Here, we want to address the question of whether immune dysfunctions detectable in the peripheral blood are also present in other form of ALS and can be used to determine disease progression and severity, as well as potential therapeutic targets.

Mining the B cell repertoire of resilient ALS patients
Alchemab Therapeutics who will utilize a novel unbiased approach to identifying new therapeutics and drug targets by evaluating convergent protective antibody responses in groups of resilient patients. The hypothesis being examined here is that ALS patients may possess auto-antibodies which provide some degree of disease resilience. Any antibodies discovered will subsequently be investigated in disease models to determine if they contribute to these patients disease resilience, with an ultimate view of developing novel therapeutics.

Clinical Development of ISGylation Biomarker for ALS
Amyotrophic Lateral Sclerosis (ALS) is a fatal disease. In Greek language, “A” means no, “Myo” means muscle, and “Trophic” means nourishment. Therefore, amyotrophic means “no muscle nourishment”. When a muscle has no nourishment, there is a loss of muscle mass, which is called muscle wasting. The word “Lateral” in ALS refers to the location of the damage in the spinal cord. This is the site where portions of the nerve cells (neurons) that signal and control the muscles are located in our body. In ALS patients, neurons in this area die, and it leads to hardening of this region, which is called “sclerosis”. Unfortunately, there is no biomarker, an indicator of ALS disease, available. Excitingly, we have found one in our laboratory, which is called “ISGylation” in the blood of ALS patients. We are requesting blood serums from ALS patients that are archived in the NAB to confirm our findings. This biomarker would help doctors to detect and manage ALS disease in patients.

Advanced Patient-Derived Microglia Assay for Preclinical and Clinical Trial Drug Validation
This project will grow brain immune cells call microglia from blood cells of people with ALS. We will use these cells to test new drugs that may be able to slow the disease in people with ALS. Our research using these cells will help determine if an individual is suited to a clinical trial for a particular type of drug. This would help align people with ALS to the correct trials. We are also examining if we can develop new markers of drug effectiveness in these cells. This will help to improve the success rate of clinical trials for drugs that target immune cells in the brain (believed to be involved in ALS). We are seeking a blood sample from people with ALS (10 different people) and these blood samples will be used to generate the brain immune cells for this research project.

Heavy metals and methylated DNA in sludge and wastewater: Population biomarkers for susceptibility to neurodegenerative diseases
Large-scale human genomic data: In Aim 3 of the proposed work, we plan to conduct an epigenome-wide association study (EWAS) study to show the importance of gene x environment interaction. We will be quantifying differential methylation signatures in human cfDNA extracted from urine and wastewater using the Methylation EPIC BeadChip (Infinium) microarray that covers over 850,000 CpG methylation sites (850K) on the human genome. Secondary human epigenome data of blood shared in databases such as the Gene Expression Omnibus (GEO) /ADNI/or ROSMAP datasets accessible by researchers will be pulled for alignment to urine and wastewater methylation results.

Biomarkers for early stage ALS detection
Clinical treatment outcomes would be enhanced if early phase detection could be achieved, perhaps through the identification of specific and selective “biomarkers” of disease stage, progression, and response to treatment. No such effective biomarkers currently exist: The result from the age and sex matched ALS patients will be compare with expected cytokines results. This study could help achieve early phase ALS detection by finding specific and selective “biomarkers” of disease stage, progression, and response to treatment. This could enhance clinical treatment in the future.

Developing low-cost technology to detect ALS Biomarkers
This project will grow brain immune cells call microglia from blood cells of people with ALS. We will use these cells to test new drugs that may be able to slow the disease in people with ALS. Our research using these cells will help determine if an individual is suited to a clinical trial for a particular type of drug. This would help align people with ALS to the correct trials. We are also examining if we can develop new markers of drug effectiveness in these cells. This will help to improve the success rate of clinical trials for drugs that target immune cells in the brain (believed to be involved in ALS). We are seeking a blood sample from people with ALS (10 different people) and these blood samples will be used to generate the brain immune cells for this research project.

To evaluate the potential biomarkers of Amyotrophic Lateral Sclerosis (ALS) using biofluids
Our research group has recently started working on neurological disorders. Our expertise is in cancer biology, molecular biology, nano formulations, drug delivery and biomarker development. The goal of this protocol is to evaluate the expression of potential Amyotrophic Lateral Sclerosis (ALS) biomarkers from biological samples derived from ALS and Non-ALS individuals. Biomarker(s) identification may help in the diagnosis of ALS as the diagnosis is the major challenge in ALS patients. ALS is not diagnosed in early stages due to the slow progression of disease or due to the presence of other neurological disorders at the same time.

ALS Subtype Biomarkers
In an earlier study using postmortem tissue, we found three major differences among ALS patients. This study will try to determine whether or not we can see similar gene expression differences in other bodily fluids. These bodily fluids could be obtained while patients are still living. We hope to obtain lists of genes measurable from CSF or blood that can predict the ALS Subtype that defines that patient.

Characterization and Identification of Biomarkers of ALS Disease Onset and Progression
We want to learn about the mechanisms of ALS disease. We want to evaluate key markers at the molecular level (including genes and proteins). We will use samples from ALS patients banked at the National ALS biorepository.

Quantification of Self-Replicating Proteins in the CSF of ALS Patients and Disease Controls
Currently, the cause of sporadic ALS is unknown, although knowledge regarding the disease has markedly increased since the identification of genetic causes of the disorder, which account for about 10% of cases. Among theories regarding sporadic disease, the idea that misfolding of cellular proteins are causative factors is a leading consideration. Recently, the technology for detecting minute amounts of proteins which, on their own, replicate has been developed. At the outset, our goal is to determine if α-synuclein is a molecular culprit in some instances of ALS. We predict that a subgroup of ALS patients will be found in which this protein is elevated. If this is the case, our finding will have implications for both the diagnosis and treatment of ALS.

ALS miRNA Biomarker
This research aims to increase our current sample size to assess if these biomarkers are a robust indicator of ALS
that could be used for clinical diagnosis. Also, to determine if plasma samples collected by different labs using different protocols are adequate to identify the miRNA biomarkers, thus increasing the chance that these biomarkers would be useful in a clinical setting.

Targeting an Immune Pathway Disrupted by Multiple ALS-Causative Genes
This research aims to increase our current sample size to assess if these biomarkers are a robust indicator of ALS
that could be used for clinical diagnosis. Also, to determine if plasma samples collected by different labs using different protocols are adequate to identify the miRNA biomarkers, thus increasing the chance that these biomarkers would be useful in a clinical setting.

Histologic investigation of extracellular innate checkpoint molecule CD47 in ALS motor neurons
In Amyotrophic Lateral Sclerosis (ALS), the underlying trigger of the catastrophic cascade of neuron functional decline, activation of immune cells, inflammation, tissue damage spread and hardening of the corticospinal tracts is unknown. CD47 is an extracellular protein used to self-identify all cells and functions by turning off the innate immune system (innate checkpoint). In this study, we will explore CD47 expression levels and location to determine if loss of surface CD47 or change in cell surface localization occurs with neural cellular dysfunction, possibly linking neural damage to innate immunity activation.

Development and validation of assays to determine target engagement in ALS clinical studies
Amyotrophic Lateral Sclerosis (ALS, Lou Gehrig’s disease) leads to changes in and ultimately the death of nerves that control muscles. As these nerves decay they undergo a number of changes. Our work is focused on developing tests that can be used to determine the effectiveness of experimental therapies for the treatment of amyotrophic lateral sclerosis by demonstrating whether or not a particular therapy reverses the disease induced changes.

The Influence of Inflammation in the Progression of ALS
This research aims to understand how inflammatory responses, especially those aided by external stimulations such as virus infection, propel the progression of ALS. To further validate their hypothesis, they hope to obtain biological information from the blood of ALS patients with specific genetic mutations.

LBT-3627: A Novel Immunomodulatory Disease-Modifying Approach to ALS Treatment
Longevity Biotech is measuring the immune function in patients with amyotrophic lateral sclerosis (ALS). De-identified patient immune cell samples will be obtained and cultured in a dish where samples will be treated with the experimental compound LBT-3627. Function will be measured using a standard suppression of proliferation assay.

RNA-Sequencing based drug discovery in ALS
The study approach combines the detailed examination of gene expression in sections from human brain using immunohistochemistry (IHC) and in situ hybridization (ISH). On identification of proteins or mRNAs expressed in and restricted to motor neurons, our discovery process moves to sub-dissection of frozen human CNS, isolation of nuclei, fluorescence activating cell sorting (FACS) and RNA sequencing with bioinformatic analysis [Xu et al, in press]. In applying this approach in the context of amyotrophic lateral sclerosis (ALS), the aim is to identify genes expressed in distinct populations of motor neurons (upper or lower) dysregulated in disease. Ideally, the targets we identify through bioinformatic analysis of sequencing data should offer the potential for therapeutic intervention through both neuroprotective mechanisms or through stimulation of key receptors. Use of high-quality human CNS samples from normal and diseased donors is therefore key to their research. The primary aim is to isolate nuclei from upper and lower motor neurons by FACs and compare gene expression by RNA sequencing between neurons and glia from healthy donors and donors with ALS.

Assessment of Unbound Free Fatty Acids in ALS Plasma
Measure profiles of unbound free fatty acids in the plasma of ALS patients to determine whether these may serve as a biomarker for the disease. Unbound FFAs were measured using fluorescently labeled mutated fatty acid binding proteins (probes). Nine FFAs comprise more than 96% of the long-chain FF in plasma. Using 20 different probes with complementary specificities for the 9 FFA we determined the concentrations of each of the 9 unbound FFA (profiles) in plasma and CSF samples obtained from the NEALS biorepository. We are requesting plasma samples from 100 ALS subjects. To the degree possible, we would like demographic information including: e.g., age, gender, race, limb vs. bulbar onset, disease duration, and drug usage.

Biomarkers in neuronal exosomes for assessment of ALS progression
To conduct a pilot, feasibility study measuring biomarkers longitudinally in neuronal exosomes isolated from the serum of a small cohort of patients with ALS. Exosomes will be isolated from the serum using the ExoQuick ULTRA kit (Systems Bioscience) followed by immunoprecipitation of neuronal exosomes as described previously (Yan et al., ACS Sensors, 2019). Following extensive washing the exosomes will be lysed and candidate biomarkers will be measured using ELISA (for protein biomarkers) or qPCR for RNA biomarkers). Patients with ALS (n = 30) from whom serum samples are available at two different time points 6-12 months apart.

Novel extracellular vesicle and molecular biomarkers of environmental exposure and disease progression in ALS
The study from the Columbia University Health Sciences will test new non-invasive ways for identifying possible toxic exposures to the brains of ALS patients. The study will explain which processes link exposures and progression of disease. The study will be the first to test whether extracellular vesicles from the central nervous system can be used as new biomarkers. These biomarkers will be measures of environmental exposures and disease progression in ALS. The researchers will test samples from persons with ALS for metals and pesticides. They will then match the exposure and patient specific transcriptomic signatures to ALS signaling pathways.

Metals analysis
There have been limited studies that measured environmental chemicals in specimens from persons with ALS. We will first focus on metals and metalloids, including mercury, cadmium, lead, manganese, arsenic, chromium, selenium, copper, and possibly others. We will examine the hypothesis that exposure to metals is associated with the getting ALS. We will use specimens that were collected as part of the ALS biorepository. We expect the sample size will be between 300 and 600. The biorepository does not include a control group. Therefore, we will use the NHANES data to obtain estimates of the distribution of metal levels for the US Population.

Genomic Analysis
DNA samples are evaluated for mutations in known ALS genes. The NeuroChip genotyping array is used. It is a low-cost, custom-designed array. It contains a tagging variant backbone of about 306,670 variants. 179,467 variants have been added. These added variants have been linked to diverse neurological diseases. These diseases include Alzheimer’s disease, Parkinson’s disease, Lewy body dementia, amyotrophic lateral sclerosis, frontotemporal dementia, progressive supranuclear palsy, corticobasal degeneration, and multiple system atrophy.

Mitochondrial DNA and Micro RNAs in Amyotrophic Lateral Sclerosis
Recent advances in the molecular genetics have increased understanding of many of the genetic/familial forms of ALS and other motor neuron diseases. Nevertheless, most ALS cases cannot be attributable to genes alone. There is much to be learned from studying environmental factors in the etiology and progression of ALS and other motor neuron diseases. Recent evidence points to a role of oxidative stress in the etiology of ALS. Recent developments in molecular epidemiology suggest that changes in mitochondrial DNA (mtDNA) and microRNAs (miRNAs) are associated with oxidative stress. The best biological specimens to measure these novel biomarkers would be the motor cortex of the brain and the spinal cord. However, these tissues are not available until after death. In this project we will compare measures of mtDNA and miRNAs in the motor cortex, spinal cord, and blood. This will help us see whether blood adequately reflects their amounts in target tissues.

Role of FUS protein in inflammation and neurodegeneration, as potentially applied to understanding the development of ALS
Several pathogenic mutations have been identified in the genome of ALS patients. Each mutation is associated with varied phenotypes. The researcher’s lab studies the function of the protein FUS. FUS has numerous variants associated with ALS. To address the cause-effect relationship between inflammation and neurodegeneration the researcher aims to study the function of FUS along with other ALS implicated proteins in the context of susceptibility to infection.

ALS risk, exposure sources, and effects on the unfolded protein response pathway.
The first phase of the proposed study will focus on comparing exposures to environmental pollutants (lead, mercury, and other heavy metals) in nails. Samples from ALS patients will come from the National ALS Biorepository. They will compare with those of controls from a large, nationally representative prospective cohort. This cohort is funded by the National Institute of Environmental Health Sciences.

Identification and Characterization of Potential Environmental Risk Factors for ALS using the ATSDR ALS Registry Cases and a Control Population.
This research will study self-reported environmental/occupational exposure to metals, pesticides, and solvents for persons with ALS and controls. It will also study life-time air pollution exposure based on where people lived. The first aim will look at a variety of air pollution data. The second aim will verify self-reported exposures to solvents and pesticides using blood measures of persistent pollutants. In aim three we will study the relationship between environmental toxicants in human samples and key biological pathways and common genes associated with developing ALS.

Targeting Ataxin-2 in Amyotrophic lateral sclerosis (ALS).
In 1996 we discovered a gene that is mutated in a rare disorder known as spinocerebellar ataxia type 2, or SCA2. The mutated gene that causes SCA2 is called ATXN2. When mutated ATXN2 gains abnormal and toxic functions. For diseases caused by such “gain of function disease genes,” it is believed that treatments that lower or eliminate their function may be therapeutic. Recent studies have shown that lowering the amount of ATXN2 also improves the survival of mice with the same abnormalities causing ALS in humans. The underlying reason for that is related to the abundance of yet another protein called staufen. Staufen is overabundant when ATXN2 is mutated. We have shown that lowering staufen works essentially as well as lowering ATXN2 for treating SCA2 mice. We also shown that staufen is overabundant in cells that are abnormal in ALS. Our hypothesis is that therapeutics that target the abundance of staufen will be as effective for treating ALS as therapeutics that target ATXN2. Moreover, targeting staufen may be an effective therapeutic approach for the treatment of other progressive neurological disorders. This research will support future work on the producing a drug that targets staufen for the treatment of ALS and potentially other neurological diseases.