VDP-132 [Funding] Mario Cordero - Multi-inflammasome inhibitors to treat aging and age-related diseases

One-liner: Development of multi-inflammasome inhibitors as a potential treatment for age-related diseases.

Longevity Dealflow WG team

• Senior Reviewers: Review requested
• Shepherd: Maria Marinova
• Other squad members: Eleanor Davies, Estefano Pinilla, Abhijit Kale, Ryan Spangler, Cassy Le, Adrian Matysek
• Sourced by: Maria Marinova

Project PI

Mario D. Cordero, PhD

Principal Investigator

Department of Molecular Biology and Biochemical Engineering, Universidad Pablo de Olavide, Seville, Spain

Simple Summary

Inflammasomes are multiprotein complexes recruited and assembled upon exposure to pathogenic microbes and host danger signals, initiating the inflammatory response and activating caspase-1. However, they are also involved in the development of inflammatory disorders and the onset of age-related diseases, such as metabolic or neurodegenerative conditions.

The Principal Investigator, Mario D. Cordero, PhD, and his team showed in their preliminary results that different inflammasomes interact in the early stages of the inflammatory response, setting the hypothesis that the formation of heterocomplexes of inflammasomes is a key regulatory pathway for for pro-inflammatory mediators such as IL-1β and IL-18.

The team will explore the molecular links between the signaling mechanisms of inflammasomes to form regulatory heterocomplexes throughout the lifetime. They will address the formation of a coordinated macromolecular self-regulatory network and its malfunction in age-associated phenotypes and during cellular senescence as a physiological source of age-associated damage. Lastly, Cordero and his team will generate an innovative set of potential therapeutic candidates that will lay the basis for novel, long-lasting effective treatments for age-related diseases.

Problem

Aging is a natural process for all living organisms based on a progressive impairment of the physiological homeostasis at the cellular, tissue, and organismal level. The biological hallmarks of aging include inflammation. “Inflammaging” refers to the progressive accumulation of pro-inflammatory mediators during biological aging creating a low- grade sterile chronic inflammation environment. Among these mediators, IL-1β and IL-18 have been extensively studied and are directly related with the formation and activation of the inflammasomes.

Inflammasomes are multiprotein complexes responsible for the regulation of caspase-1 activation and the induction of inflammation in response to infectious microbes and host danger signals. However, inflammasomes activation in microglial cells and macrophages is associated with aging and age-related neurodegenerative diseases.

It is therefore paramount to not only understand the physiological function of inflammasomes, but to also explore ways to regulate it in order to develop short and long-term personalized treatments.

Opportunity

The primary objective is the development of therapeutics for age-related diseases through an in-depth study of inflammasomes, crucial agents in immunity and inflammation. To achieve this goal, the present project aims to study how the inflammasomes form heterocomplexes taking advantage of similar domains to participate in the aging process to trigger age-related diseases, and acting as sensors of danger signals associated to aging.

The investigator propose to reach these goals by using a multi-disciplinary approach including cell culture, molecular biology, structural biology, biophysics and animal models to specifically address the following objectives:

WP1.- Characterization of the heterocomplexes involved in life and aging.

WP2.- Mechanism of interaction of the inflammasome to participate in the formation of heterocomplex structures.

WP3.- Design of novel pharmacological approaches to modulate superinflammasomes activity.

The research fills a critical gap in the current market, as there is a lack of comprehensive solutions targeting the complexities of inflammaging.

Other research predominantly focus on the development of inhibitors targeting individual inflammasomes. However, this approach often overlooks the compensatory mechanisms that can arise, diminishing long-term efficacy. With this project, the team is introducing a novel therapeutic target that can prevent age-related diseases by pioneering a new approach in the management of inflammaging.

Experimental plan and budget

Research Plan

Objective 1. Characterization of the inflammasomes macro-complexes involved in life and aging.

The investigator’s preliminary data showed an activation of NLRP1 and NLRP3 in an irradiated cellular model of senescence and animal model of accelerated aging (Figure 1). And other inflammasomes have been shown to be activated during aging such as NLRC4 or AIM-2. All this data opens the door to the involvement of multiple inflammasomes in various stages of aging and senescence with interaction between them.

Figure 1. Inflammasomes expression during aging and senescence

Objective 2. Human validation of the inflammasome heterocomplexes

In numerous human diseases where the inflammasome has been studied, multiple papers provide evidence of various activated inflammasomes. Consequently, several cardiovascular, metabolic, and neurodegenerative diseases have demonstrated elevated levels of NLRP1, NLRP3, NLRC4, or AIM2 inflammasome complexes. The investigator and his team aim to validate findings from other objectives using human samples obtained from diverse diseases.

Objective 3. Design of novel pharmacological approaches to modulate heterocomplexes

The investigator will use the structural data available on the different individual inflammasomes to perform a rational-driven search for novel modulators of heterocomplexes.

The compound will also be evaluated in vivo models of aged mice, accelerated aging model (progeroid mice) and a gain of function NLRP3 mutant associated with neonatal onset multisystem inflammatory disease (NOMID) in mice. The investigator will analyse survival (Kaplan–Meier survival curves) bodyweight, food and water consumption, cardiometabolic profile (electrocardiographic examination, glucose tolerance and insulin response, Leptin, adiponectin and serum biomarkers) and inflammatory profile (Cytokine Array / Chemokine Array 42-Plex with IL-18 (HD42)), histological of liver, heart, kidney, lung, muscle, ovary and skin from male and female mice.

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Moreover, Cordero’s laboratory has developed a compound with the potential to inhibit both NLRP1 and NLRP3, which are two major inflammasomes involved in distinct inflammatory pathways. Simultaneously targeting both could offer a broader spectrum of action, impacting various inflammatory conditions or diseases. Preliminary data indicate that NEK7 is an essential protein in the activation of both NLRP1 and NLRP3 (Figure 2). Cordero’s initial compound, C3, demonstrates significant effects in inhibiting the interaction of NEK7 with NLRP1 and NLRP3.

Figure 2. Experimental confirmation of an inflammasome heterocomplex by Native PAGE and NMR.

Budget

Personnel

Postdoctoral researcher: 25.500/year (2 years)

Total (2 years): USD$ 51 000

The investigator requests hiring of a postdoctoral researcher to work on the objectives of the project and give stability to the research team of Dr. Mario D. Cordero. The rest of the requested budget is for consumables, reagents and animal facility and mice cost, central services, publication expenses and registration fees for congresses and trips for the dissemination of the results of the project.

|Consumables|Year 1|Year 2| | --- | --- | --- | |Animal costs (mice, animal facility)|8 800|8 800| |Chemical synthesis||8 000| |Antibodies|6 000|4 500| |Reagents|10 000|9 000| |Elisa Kits|3 500|3 000| |X-ray facility|800|1 000| |Microscopy facilities|3 500|3 000| |Western blots assays|6 000|5 000| |Statistical assistance||1 500| |Publications|4 000|4 000| |Travel allowances| |Registration, accommodation, transport|5 000|5 000| |Sub-total|47 600|52 800| |Sub-total execution costs and travel allowance:|USD$ 100 400| |IN TOTAL|USD$ 151 400|

VitaDAO Funding Terms

Sponsored Development Agreement or Decentralised Tech Transfer model (reference: VDP-45) with terms to be agreed with TTO. Discussion ongoing.

Relevance to longevity

This project is highly relevant to longevity due to its focus on inflammasomes, which are multiprotein complexes involved in both the inflammatory response and the onset of age-related diseases like metabolic and neurodegenerative conditions. The Principal Investigator, Mario D. Cordero, and his team aim to explore how different inflammasomes interact in early inflammatory stages and hypothesize that their formation into heterocomplexes is a crucial regulatory pathway for these proteins.

Understanding these molecular links and the formation of regulatory heterocomplexes throughout a lifespan could uncover insights into age-related damage and cellular senescence. By investigating the malfunction of these complexes in age-associated phenotypes, the team hopes to generate potential therapeutic candidates for treating age-related diseases more effectively.

Team

Dr. Mario D. Cordero, Principal Investigator

Dr. Antonio Astorza, Postdoctoral Researcher

Dr. Juan Miguel Suarez, Postdoctoral Researcher

Inés Muela, Predoctoral Student

Juan A. Pedregal, Agent of the Intellectual Property office of the Universidad Pablo de Olvide

Strengths

Innovative Approach: The focus on studying inflammasome heterocomplexes during aging is innovative, offering a unique perspective on the inflammatory processes associated with aging.

Preliminary Compound Development: Having a family of compounds with inhibitory potential against a heterocomplex formed by NLRP1/NLRP3 provides a strong starting point for drug development.

Experienced Research Team: The expertise of the Principal Investigator and his team, backed by previous research demonstrating the roles of inflammasomes in various aspects of aging, serves as a strong foundation for this project.

Risks

Complexity of Inflammasome Interactions: Understanding the interactions between different inflammasomes can be highly complex. Validating specific heterocomplexes associated with aging and diseases might face challenges due to the intricate nature of these molecular interactions.

Translation to Therapeutics: While the preliminary compounds show promise, translating them into effective therapeutics for human use might face hurdles such as toxicity, off-target effects, or limited efficacy in vivo.

Long-Term Effects and Safety: Given that treatments for age-related diseases might require long-term administration, ensuring the safety and efficacy of inhibitors over extended periods, especially in aging populations, could pose challenges.