NIH/NCI 361: Highly Innovative Tools for Quantifying Redox Effector Dynamics in Cancer

Fast-Track proposals will not be accepted.

Direct to Phase II will not be accepted.

Number of anticipated awards: 2-4

Budget (total costs, per award):

Phase I: up to $225,000 for up to 9 months

Phase II: up to $1,500,000 for up to 2 years

PROPOSALS THAT EXCEED THE BUDGET OR PROJECT DURATION LISTED ABOVE MAY NOT BE FUNDED.

Summary

The generation and dynamic interplay of redox effector molecules (e.g., oxygen, free radicals, peroxides, nitrogen oxides, and hydrogen sulfide) are fundamental features underlying the genomic, structural, metabolic and functional alterations observed in cancers. Alterations in redox balance impact all phases of disease including carcinogenesis, disease progression, response to treatment and prevention. For example, the DNA damaging effects of free radicals can mutagenize key oncogenic sites. Redox imbalances occur by abnormalities commonly associated with cancers including mutations in p53, myc and ras pathways. Redox effectors operate to modify protein function at the post-translational level, which plays a significant mechanistic role in the phenotypic plasticity cancer cells demonstrate in the face of oxidative and reductive (hypoxia) stresses. Redox tone is a key regulator of the self-renewal properties of stem-like cancer cells, which has been shown to contribute to tumor resistance to current therapies.

Progress in the cancer biology and pre-clinical space has been limited by the lack of tools that can accurately measure redox parameters in animal models with sufficient spatio-temporal resolution and minimal perturbation of the system. NCI seeks input from the small business community to develop and optimize a new generation of quantitative and specific technologies that will enable and accelerate basic research aimed at understanding basic redox effector mechanisms and the roles they play in the cellular adaptations and complex biology of tumors.

Supporting the development of these technologies will allow researchers to validate and benchmark data obtained across different 3D cell culture platforms and pre-clinical animal model systems with the goal of accurately mimicking tumor environments experienced by patients with cancer. Moreover, an enhanced ability to screen, manipulate, or analyze redox dynamics is an invaluable index in the evaluation of cancer cell-tumor responses to therapeutic interventions in the critical pre-clinical testing phase. These redox data have potential to significantly improve our understanding of tumor biology and ability to better predict treatment responses and long-term efficacy when translated into patients.

Project Goals

There is an unmet need in basic cancer research for probes or technologies that can better measure, characterize, profile, or resolve the spatiotemporal dynamics of redox effectors at the subcellular to cellular levels.  Genomic profiles, for instance, cannot capture post-translational redox regulation that occurs with changes in the tumor microenvironment.   Redox probes have been traditionally reliant on organic dyes that experience spectral shifts with redox.  The current state of the art is genetically encoded redox indicators that couple redox responsive enzyme motifs with indicator proteins.  These genetically engineered redox probes have improved response kinetics, but may have limited optical qualities.  Given the critical role played by redox effectors, developing a range of new tools will help us better understand how redox effectors regulate cell phenotypes in functional tumor populations.

The goal of this FOA is to develop quantitative tools to measure redox dynamics in biological systems.  Ideally, probes or biosensor tools should be minimally invasive as to not significantly perturb the system.  The technical approach should: (1) allow for in vivo measurements of redox effector spatiotemporal dynamics; and-or (2) be useable in high throughput systems, for example to allow the screening of cellular response to experimental perturbations, such as exposure to cytotoxic agents.  The long term goal is that the technologies developed through this contract can help validate whether data gathered in model experimental systems faithfully represents the redox dynamics of human tumors.

Technologies that have the potential for in vivo use, especially those with potential clinical applications in the long term will be of particular interest, but methods that will be restricted to pre-clinical research applications are also of interest.

To successfully meet this goal, offerors shall develop a technology for the minimally to non-invasive measurement of one or more redox effectors, including but not limited to oxygen, free radicals, reactive oxygen species, peroxides, nitrogen oxides, and hydrogen sulfide.  Phase I studies should focus on developing the technology and demonstrating proof of concept in an in vitro system. Phase II studies further refine the technology and demonstrate the use of the technology to measure redox effectors.  Offerors shall justify their choice of approach with respect to the scientific utility and commercial potential, and specify quantitative milestones that can be used to evaluate the success of the technology being developed.

It is anticipated that offerors shall develop a probe or similar agent that facilitates the measurement of redox effectors by one or more imaging modalities; however, offerors are not restricted to any particular technical approach and label or probe free approaches that can meet the requirements of this contract are welcome.

Offerors are not restricted to any particular technical approach and can propose resource and tool development that incorporates high-risk/high-impact technologies.  Examples can include, but are not

limited to:

• Redox probes that provide significant advances in sensitivity, selectivity, ratiometric capability, or resolution in reporting the spatial concentration gradients and temporal dynamics of redox effectors at the subcellular, cellular and/or tissue compartment levels.
• Genetically encoded redox biosensors that are expressed in a cell or tissue selective manner in small animal models of cancer for interrogation by non-invasive to minimally invasive imaging modalities.
• Biology-inspired redox sensors (e.g., based on bacterial chemosensors) that through synthetic biology techniques are genetically encoded for expression in a cell or tissue selective manner.
• Nanotechnology scaffolds multiplexed with sensors that permit functional parallel profile analyses of a combination of redox effectors (i.e., oxygen, nitric oxide, hydrogen peroxide, superoxide) and/or related species (e.g., proton, glutathione, ascorbate) across both time and space at the subcellular, cellular and/or tissue compartment levels.
• Instrumentation that enables label-free quantitative measurements of redox-related spatiotemporal dynamics in cancer cells and/or tumors (e.g., Raman spectroscopy-based microscopy, super resolution microscopy).

Technologies that have the potential for in vivo use, especially those with potential clinical applications in the long term will be of particular interest.  However, Offerors with technologies that will advance pre-clinical or basic cancer research applications are also of high interest.

Phase I Activities and Deliverables

• Identify and justify development of a sensing tool or probe for specific redox effector species from both a cancer biology and commercial perspective.
• Offerors shall describe the current state of the art technologies for sensing and measuring the redox effector being addressed by their proposal, and outline the advantages that their approach will offer.
• Develop and characterize a redox probe, biosensor or detection platform. Offerors shall specify quantitative milestones that can be used to evaluate the success of the technology being developed, and justify these milestones from the viewpoint of both scientific utility and commercial value.
• Develop an assay or system that demonstrates proof-of-concept testing and benchmarking of specificity and sensitivity parameters of the agent or system for a range of redox effector species (e.g., oxygen, free radicals, hydrogen peroxide, nitric oxide, hydrogen sulfide, NAD/NADH, GSH/GSSG).
• For each redox effector or parameter, a technical description of methodology for each assessment shall be provided that includes how each measurement is calibrated.  If measurements are collected serially, the rationale for the order of measurements shall be specified.
• Demonstrate feasibility to sense, interrogate, detect or resolve the spatiotemporal dynamics of redox effector species in live cells or animal model, ideally with a minimally invasive perturbation of the system.
• Provide NCI with proof-of-concept assay SOP. 

Phase II Activities and Deliverables

The goal of the Phase II product is an optimized commercial resource, reagent, kit or device that can allow researchers to measure the relevant redox effector molecules in their laboratory.  Decisions for continued project development into Phase II will be based on probes, biosensors, assays or systems that:

• Can demonstrate reliability and robustness.  Offerors shall provide a technical evaluation and quality assurance plan with specific detail on shelf life, best practices for use, equipment required for use.
• Can be scaled up at a price point that is compatible with market success and widespread adoption by the basic research community.
• Have potential to benchmark data obtained across different cancer model systems. 

Deliverables for the Phase II projects are:

• Scaled up synthesis or manufacture of agents, chemicals, device, or products necessary.
• Design and implement quality assurance controls and assays to validate production.
• Validate scaled up device, chemical or product. Offeror shall demonstrate the utility, reliability and sensitivity of their device, chemical or product across in vitro and/or in vivo models relevant to cancer research.
• Refine SOPs to allow for user friendly implementation of technology by the target market for the agents, chemicals, device, or products.