My name is Evangelos Kiskinis. I'm an Assistant Professor of Neurology and Neuroscience at Northwestern University, Feinberg School of Medicine in Chicago, Illinois.
The overall objective of our work is to try and understand how rare genetic mutations that have been associated with neurological diseases such as amyotrophic lateral sclerosis (ALS), or pediatric forms of epilepsies, impact the function of human neuronal subtypes and lead to their eventual degeneration. We believe that if we understand those processes, we'll be able to design rational therapeutic approaches that will allow us to slow down or even reverse these pathological disease processes that are associated with these patients.
The primary model system that we use in my lab is patient-specific induced pluripotent stem cells. So, we typically recruit a patient, either an ALS patient or a child that's been diagnosed with pediatric epilepsy through one of our neurology clinics here at Northwestern and we isolate a somatic cell type from that individual patient. These days, that's typically a few millimeters of their blood.
We then reprogram those blood cells to generate patient-specific induced pluripotent stem cells, which we can then differentiate into distinct neuronal subtypes that are affected in these particular diseases.
So, in the case of ALS, these are spinal cord motor neurons or cortical motor neurons. And in the case of pediatric epilepsy, there is different neuronal cell types that are found within the cortex. We then study the functional properties of these cells by employing a range of different assays from omics analysis to electrophysiological analysis to define how the mutations affect the ability of these cells to function and to also identify particular pathological features that are also characteristics of these patients.
Gene editing has been a transformative tool for us because it has allowed us to test the contribution of particular mutations toward pathological phenotypes that we identify in the iPSC-derived neuronal systems. So, in the case where we know a particular mutation that has been associated with a particular patient, we use CRISPR-Cas9 to correct that mutation and generate what we refer to as an isogenic control stem cell line.
Then we take the patient stem cell line and the corresponding isogenic control, and we differentiate these into different neural subtypes in parallel. And by studying the subtypes in parallel, we're able to attribute particular phenotypes to that mutation. We can also perform the opposite experiment to test the sufficiency of a specific mutation where we take an iPSC line derived from a healthy control individual and we introduce a mutation of interest.
And if we see that that mutation is able to recapitulate a particular phenotype, then we conclude that the mutation is sufficient in generating that pathology. The most exciting developments associated with CRISPR-Cas9 or base editing technologies are the therapeutic approaches that are already clinical trials looking at ex vivo gene editing for particular blood diseases.
There's also a lot of exciting pre-clinical work in various in vivo model systems using gene editing technologies or base editing technologies that has shown that you can really correct mutations in DNA in various animal models.
I think the application of these technologies will be transformative in our ability to treat individuals that are suffering from these debilitating genetic diseases.
