One can never know where research will lead. It’s fun to trace milestone achievements and with hindsight, identify the tipping point. There is a theory, Black Swan Theory, that is a metaphor that describes an event that comes as a surprise, has a major effect, and is often obvious with the benefit of hindsight. It is perhaps why great discoveries are simultaneously “discovered” by several scientists in different areas of the world, because the idea’s time has come. Foundational science supported a leap forward. History is full of such discoveries that were hinted, and then exposed perhaps 100 years later, because the right mind’s mused and connected the dots in a different, but obvious pattern.
The field of biotechnology that consumes my time now started the year I was born. Coincidently Thomas Brock earned a PhD in botany that year. He served in WW II and was able to continue his education on the G.I. Bill. In the mid-sixties, I was learning to ride horses while he was conducting field research on microbes in Yellowstone National park. Fortuitously, that year he and his student isolated a pink bacteria, they called T. aquaticus, from Yellowstone’s Mushroom Hot Spring. This was a tipping point.
By the time I had my MS and was working in a biochemistry laboratory, Brocks team had isolated DNA polymerase from T. aquaticus. The ability of T. aquaticus to tolerate and thrive in the hot springs was due in part to its DNA polymerase enzyme. In a few short years the enzyme, that worked at 160 F, would be exploited speeding up DNA reactions in test tubes. Selecting a difficult-to-find sequence of nucleic acids from a mix of many samples is accomplished by initiating a chain reaction, polymerase chain reaction or PCR, not conceptualized in 1976.
I left research for veterinary medicine at the time Kary Mullis was musing about a method to find point mutations in human DNA. I just moved to my farm in 1985 and that was when Mullis combined the thermophilic DNA polymerase in a chain reaction by temperature cycling and thereby revolutionized the field of biochemistry into molecular biology. This was a black swan event.
Mullis thought of making 2 opposing, complimentary primers, followed by multiple replication cycles, to amplify the difficult-to-find sequence. It was tedious. But by 1983, T. aquaticus provided a heat stable polymerase to streamline the process into PCR. He received the Nobel Prize in 1993. By the time I returned to biochemistry research in 1999, the field of biotechnology was well underway, in part due to PCR.
My interest has shifted from sarcocystosis in horses to amyotrophic lateral sclerosis, ALS, in the last two years. ALS was nationalized with the ice bucket challenge in 2014 by pro golfer Chris Kennedy and Pete Frates. Frates was diagnosed with ALS when he was 27. He died at 34.
ALS, or Lou Gehrig’s disease, is a degenerative motor neuron disease that results in loss of neurons that control voluntary muscles. Genetic and viral causes are thought to be involved in 95% of cases, while the remaining cases are inherited from a person’s parents. The disease was first described in 1824 and then connected to symptoms in 1869. Lou Gehrig was diagnosed in 1939 and, most famously, Stephen Hawking in 1963.
The first gene associated with ALS was discovered in 1993 and the first animal model, the transgenic SOD1 mouse, in 1994. Yet, today there is no cure and current goals are to improve symptoms. There are a few recently proposed biomarkers designed to assess treatments. We were told by a leading ALS researcher/clinician that if he knew the disease process, he could treat his patients. Until doctors have good biomarkers to pinpoint the pathology in a patient, there is little hope of treating ALS. And patients die.
There are several, perhaps ten or more, pathologies associated with ALS and researchers are working on each aspect of the spectrum of diseases that define ALS. A short list of pathologies includes nucleocytoplasmic transports defects, RNA metabolism dystrophies, toxic protein aggregation, dysfunctional DNA repair, mitochondrial dysfunction/oxidative stress, oligodendrocyte dysfunction, microglial dysfunction, axonal transport defects, vesicle transport defects and glutamate excitotoxicity at the neuromuscular junction. Inflammation is a key aspect of some presentations.
ALS is the most common motor neuron disease in adults and the third most common neurodegenerative disease (Alzheimer’s disease and Parkinson’s disease are more common). Riluzole was the first FDA-approved treatment with slight efficacy to increase life span by 3-4 months, it was licensed in 1995. Edaravone was approved in 2017 and helps manage ALS, a new oral formulation started clinical trials in 2020. The expectation of Edaravone is to extend life a few months.
Work on basic science after the molecular biology revolution identified pathological pathways involved in ALS. New treatment approaches involve turning genes on-- or off, are conceptualized and reasonable approaches are in clinical trials. ALS researchers are poised to find that translational piece of technology or an idea that will revolutionize treatment of this disease--it is frustrating to think the missing piece is before us, yet unseen. It might be the musings of one person to find something great. But more likely, it will be a group effort. With a foundation in basic science and a spark of an idea we can find an effective treatment. ALS is poised for the idea that will be a black swan event.
Our efforts in the last year have connected 15 researchers across the United States working on novel ideas in:
- Identifying markers for stressed tissue from adipose derived stem cells (ASC’s)
- Investigating immune checkpoint cell signaling
- In vitro analysis of perivascular stem cell repair of cell damage
- Testing the effect of combinations of therapeutics in SOD1 mouse models of ALS
- Evaluate the components of conditioned media from ASC’s
- Develop cGMP secretome for a feasibility study in ALS patients
- Develop drug analogs to increase effectiveness and decrease toxicity in ALS therapies
- Analysis of DNA and RNA to define ALS phenotypes
- Investigate pathogenic mechanisms of protein kinases in progressive neurodegenerative disorders
- Investigate axonal transport and phosphokinase signaling in ALS
- Testing potential action of compounds on axonal transport and phosphokinase signaling in vitro
- Clinical study investigating metabolomics/miRNA study in ALS/control patients
- Detection of serum analytes found in neurodegenerative diseases by electron transfer
- Identify dipeptide nucleic acids by size (iPSC or PBMC)
- Design and fabricate multiplex chip wax-on-paper sensors for proteins and nucleic acids of interest.
Hopefully the synergy in our group will bring something transformative to those that are patiently waiting, those afflicted with ALS.