Bosco Lab

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Research

Research Overview

Our lab is investigating the pathogenic mechanisms of ALS-associated proteins SOD1, FUS/TLS, profilin-1 and TDP-43. ALS (amyotrophic lateral sclerosis), also known as Lou Gehrig’s disease, is a fatal neurodegenerative disorder that targets motor neurons. Motor neuron death culminates in paralysis and eventual death, usually 2-3 years after symptom onset. To date, there is no cure or effective therapies for ALS. Our ultimate goal is to translate our basic-research findings into therapies for this devastating disease. We use a multidisciplinary approach involving biochemistry, cell biology (including iPS cell technology), biophysics and in vivo model systems for our investigations.

Protein misfolding in ALS

Many neurodegenerative disease-associated proteins become misfolded as a consequence of genetic mutations and/or altered post-translational modifications. Misfolded proteins exert toxic functions by impairing the quality control systems within the cell and by engaging in aberrant protein interactions. Moreover, misfolded proteins self-associate and form pathological aggregates observed in post-mortem CNS tissues from individuals with various neurodegenerative diseases. Our lab is focused on defining the misfolded conformation(s) associated with pathogenic forms of SOD1 (Rotunno, JBC, 2014), profilin-1 (Boopathy, PNAS, 2015) and FUS/TLS (Sama, Sci Rep, 2017) using various biochemical and biophysical methods.  We then use this information to develop small-molecule screens to identify compounds that can  restore the protein’s normal                                                                               conformation.

Cellular stress response in disease

We showed that endogenous FUS translocates from the nucleus to the cytoplasm under hyperosmolar stress (Sama, J Cell Physiol, 2013; *cover story). We are currently investigating the effect of different stressors on disease-associated RNA-binding proteins, including FUS (Tischbein, in preparation and Baron, in preparation). Our central hypothesis is that these disease-associated RNA-binding proteins play a normal role in stress response, but that role becomes compromised or altered in disease.

Modeling ALS with human induced pluripotent stem cells (iPSCs)

We are currently studying mechanisms of ALS with patient derived human iPSCs. Mutations are corrected or introduced using CRISPR/Cas9, resulting in isogenic lines for our studies. Currently we culture lines harboring ALS-linked FUS and profilin-1 (PFN1) mutations, and differentiate these into different cell types including motor neurons and microglia for our investigations.

Stress granule biology in vivo

We showed that ALS-linked FUS incorporates into stress granules under various conditions of cellular stress (Bosco, Hum Mol Genet, 2010), and that incorporation of ALS-FUS into stress granules alters the functional properties of these structures (Baron, Mol Neurodegener, 2013). Recently, we identified the proteins that mutant FUS interacts with when forming stress granules (Yonutas, in preparation). Our current efforts are focused on studying SG-formation in vivo.