From Viral Invasion to Cellular Homeostasis: Dissecting the Impact of HSV-1 on Centrosome and Cilia Architecture and Regulation

Centrosome elimination mechanisms remain largely underexplored.-why Our preliminary research suggests HSV-1 induces CCC destruction, offering a novel angle to explore both CCC homeostasis and HSV-1 infection dynamics (Fig. 2 & 3). Our investigations, alongside recent studies, underscore the critical roles various centriole components play in maintaining stability (REF; Fig 1C, see state-of-the-artstate-of-the art section). The virus may target specific, crucial elements like ANA1/CEP295 or the PCM, both proven essential for CCC integrity (Fig. 1C), or it might exert a broader impact, affecting the CCC overall structure through mechanisms such as autophagy or widespread protein degradation. Its influence could be direct, through viral proteins like ICP0 (Fig. 4) that mediate protein degradation (REF), or indirect, by altering cellular processes such as proteostasis. Our GENERAL OBJECTIVE is to unravel those complex interactions, which will not only shed light on centriole stability mechanisms but also reveal how viral activity can alter cellular function, potentially identifying new viral vulnerabilities. Our objectives aim to:
• SPECIFIC OBJECTIVE 1 (Task 1): Unravel how HSV1 disrupts CCC architecture and its cellular consequences. We'll employ high spatio-temporal resolution and quantitative analyses to assess infection impacts on CCC and cellular health. Outcome: This approach will generate new hypotheses about key CCC components involved and how these alterations influence both the cell and infection dynamics.
• SPECIFIC OBJECTIVE 2 (Task 2): Unravel the cellular consequences, in particular for infection of HSV1-mediated disruption of the CCC. Understand how CCC disruption is important for infection, and whether there are differences encountered in different cell types that react differently to HSV-1 infection. Outcome: This task will lead to a better understanding of the cellular and infection context that surrounds CCC destruction.
• SPECIFIC OBJECTIVE 3 (Tasks 3 and 4): Identify the mechanisms behind CCC disruption by viral infection. Through both targeted and novel screening approaches, we aim to uncover and then meticulously examine these mechanisms, leveraging resources from both the centrosome/cilia and viral research labs. Outcome: We'll clarify how HSV1 impacts CCC, potentially identifying targets for manipulating these mechanisms in the context of infection.
• SPECIFIC OBJECTIVE 3 (Task 5): Validate the identified mechanisms in vivo. Utilizing skin-on-a-chip models that mimic HSV-1's primary infection sites, we will explore the implications of these findings. Outcome: This will not only confirm the in vitro observations but also provide insights into how these mechanisms operate within a living organism.
• SPECIFIC OBJECTIVE 4 (Task 6): The mechanisms through which HSV-1 interacts with centrosomes, influencing their structure and function during the viral lytic cycle, might also play a crucial role in the process of viral reactivation from latency. Outcome: By understanding these interactions, we aim to uncover whether the same or similar molecular pathways and cellular responses are involved in initiating HSV-1 reactivation, providing new insights into the virus's life cycle and potential therapeutic targets for preventing recurrent infections.
• SPECIFIC OBJECTIVE 54 (Task 7): Extend our understanding of centriole destruction mechanisms to physiological models, such as oogenesis and muscle development. Outcome: This broader examination will help ascertain the generality of the uncovered mechanisms in the regulation of the cytoskeleton.

OVERALL OUTCOME AND FEASIBILITY: These objectives collectively aim to advance our understanding of how HSV-1 interacts with centriolar structures, with the ultimate goal of uncovering new insights into both viral pathogenesis and the maintenance cellular homeostasis.