Investigating the microtubule dynamics control of KANK1 in human iPSC-derived neurons

Objectives

The main objective of this project is to understand how the protein KANK1 controls the microtubule dynamics in human iPSC-derived neurons. To this end, the student will combine several approaches including live microscopy imaging, micropatterning and biochemistry on cortical neurons derived from human iPSC.

Abstract

Amyotrophic lateral sclerosis (ALS) represents a rapidly progressing neurodegenerative disease and is characterized by a degeneration of the motor neurons. A recent study which combined machine learning approach with data from induced pluripotent stem cells-derived motor neurons from patient with ALS, revealed that KANK1 was associated with the highest number of ALS-associated genetic variants. However, the role of KANK1 in neurons still remains to elucidate. KANK1 is a protein known to link microtubule tips with the major component of integrin-mediated focal adhesion, talin. Our hypothesis is that KANK1 interaction with microtubules in the vicinity of integrin adhesions is critical and an impairment of these interactions can affect the balance between stable and dynamic microtubules, found altered in ALS. By combining a broad range of approaches, the student will (i) characterize the integrin-mediated adhesions of iPSC-derived neurons and (ii) will address in this context, the role of KANK1 in the microtubule dynamics.

Methods

Cell culture and iPSCs differentiation, Live microscopy imaging (TIRF, FRAP, Confocal imaging,) Immunofluorescence, Micropatterning, Biochemistry (Western Blot, Pull-down), Data analysis (FIJI).

References

• Aureille J. et al. (2023) Focal adhesions are controlled by microtubules through local contractility regulation. biorxiv.
• Peris et al. (2022) Tubulin tyrosination regulates synaptic function and is disrupted in Alzheimer's disease. Brain.
• Rafiq NBM et al. (2019) A mechano-signalling network linking microtubules, myosin IIA filaments and integrin-based adhesions Nat. Material.

Contacts

Julien Aureille (Research fellow)
Email : julien.aureilleuniv-grenoble-alpes.fr
Phone : 04 56 52 05 48