Research team
Expertise
My research is currently focused on the role of the Kv7.2 channel in Kv7-related epileptic and developmental encephalopathies.This research involves culturing human pluripotent stem cells (hPSCs) and differentiating them into astrocytes, excitatory cortical neurons or inhibitory neurons. Spontaneous, electrical and drug-induced neuronal activity is recorded with a high-density microelectrode array (MEA). The results of the MEA experiments are analysed with statistical programmes in GraphPad Prism and R script. Molecular biology techniques and RNAseq are part of my current project. During my PhD and first year as a post-doctoral researcher, I focused on the identification of novel Kv7 channels openers to treat Kv7-related disorders and epilepsy. I used secondary cell lines to create Kv7 channel expression models and use them in in vitro fluorometric and colorimetric drug screening assays; I used the whole-cell patch-clamp technique in heterologous expression systems to study the effect of drugs on Kv7 channels. Moreover, I performed the isolation of primary cells (endothelial and astrocytic cells as well as microvessels) from mouse/rat brains. My researcher expertise also includes scientific writing (papers and grants), presenting to a scientific audience, communication to a non-scientific audience a lay version of my work, and mentoring undergraduate students.
Investigating the Pathophysiological Role of Astrocytic KCNQ2 Channels Using Human iPSC Models.
Abstract
The Kv7 channels, encoded by KCNQ genes, are potassium channels playing a critical role in regulating neuronal excitability and stabilizing neuronal membrane potential. Within this family, the KCNQ2 channel, highly expressed in neurons, has been extensively studied due to its association with epileptic disorders. Mutations resulting in loss or gain of function (LoF or GoF) of KCNQ2 channels can lead to a spectrum of conditions, from self-limited neonatal epilepsy to severe developmental and epileptic encephalopathy (KCNQ2-DEE). KCNQ2-DEE pathophysiological mechanisms remain largely unknown; beyond epileptic seizures, patients with KCNQ2-DEE exhibit profound neurodevelopmental impairments for which there are currently no adequate treatment options. Recent evidence has revealed that KCNQ2 is also expressed in astrocytes, although its function in these glial cells is still poorly understood. Astrocytes are critical regulators of brain homeostasis, ensuring potassium and glutamate buffering, providing metabolic support to neurons, and contributing to synaptic modulation. Increasing evidence indicates that astrocytic dysfunction plays a key role in the pathogenesis of developmental and epileptic disorders. Based on these observations, I hypothesize that pathogenic KCNQ2 mutations disrupt essential astrocytic functions, thereby altering the neuronal microenvironment and contributing to the neurodevelopmental and epileptic features of KCNQ2-DEE. To test this hypothesis, I will generate human induced pluripotent stem cell (iPSC)-derived astrocytes from KCNQ2-DEE patients carrying pathogenic LoF and GoF variants, using an optimized differentiation protocol recently established in our laboratory. I will comprehensively characterize these patient-derived astrocytes by assessing key physiological processes that underlie astrocyte-to-neuron support, including calcium signaling, neurotransmitter uptake, and metabolic activity. In parallel, transcriptomic analyses will be performed to identify molecular pathways altered by KCNQ2 dysfunction, highlighting potential targets for therapeutic intervention. This BOF SRG project will, for the first time, elucidate the physiological and pathological roles of the KCNQ2 potassium channel in human astrocytes. By integrating patient-derived models and functional assays, the proposed research will address a major gap in understanding the pathophysiology of KCNQ2-DEE. The expected findings will not only advance our fundamental knowledge of this severe disorder but may also open novel therapeutic avenues aimed at restoring astrocyte function and improving outcomes for KCNQ2-DEE patients.Researcher(s)
- Promoter: Carotenuto Lidia
Research team(s)
Project type(s)
- Research Project