Development and Validation of Harmonized Methodologies to Measure NAD+ and Related Metabolites in Clinical Trials (U01 Clinical Trial Required) | RFA AG 24 039

FAQs: NIH RFA-AG-24-039 (U01) - Harmonized Methods to Measure NAD+ and Related Metabolites in Clinical Trials

What is the title and identifier of this NIH funding opportunity?

The opportunity is NIH Notice of Funding Opportunity (NOFO) RFA-AG-24-039 titled "Development and Validation of Harmonized Methodologies to Measure NAD+ and Related Metabolites in Clinical Trials (U01 Clinical Trial Required)."

What is the main problem this program is trying to solve?

The program targets a practical barrier in NAD+ research: different labs and clinical studies often measure NAD+ and related metabolites using different methods. Those differences can create method-driven variability that makes results difficult to compare across clinical trials, study sites, and sample types. The goal is to reduce that variability by creating standardized, harmonized methodologies that work reliably in human clinical research.

What is the central goal of the NOFO?

The central goal is to create, rigorously test, and validate standardized, harmonized protocols for measuring NAD+ and its related metabolites in humans so results are reliable, reproducible, and comparable across studies.

Is the focus only on developing an assay?

No. The NOFO emphasizes the entire measurement pipeline, not just the analytical assay. That includes how samples are collected and handled, how samples are stored, how metabolites are extracted, how assays are calibrated, and how results are reported so data can be meaningfully compared across studies.

What kinds of methodologies are applicants expected to develop and validate?

Applicants are expected to develop and validate protocols for quantifying NAD and related metabolites in humans, with careful attention to the pre-analytical and analytical steps that can distort measurements and reduce comparability across trials.

What are "pre-analytical variables" in the context of this NOFO?

Pre-analytical variables are factors that can affect measurements before the assay is run, such as how and when samples are collected, which anticoagulants are used, how long processing is delayed, temperature control during handling, and how samples are stored. The NOFO highlights these variables because they can substantially influence NAD+ and metabolite stability and measured values.

What sample collection factors does the NOFO call out as important to standardize?

Examples specifically noted include timing of collection, choice of anticoagulants, processing delays, and temperature control. The intent is to standardize collection procedures so measured metabolite levels are not artifacts of different handling practices.

What sample storage factors are emphasized?

The NOFO highlights standardizing storage conditions such as freezing approaches, tolerance to freeze-thaw cycles, storage duration, and use of stabilizing agents if applicable. These details matter because storage can change metabolite integrity and comparability across sites.

Does the NOFO address extraction methods?

Yes. It explicitly expects attention to extraction methods for different biospecimen types, recognizing that different sample types can require different extraction workflows and can introduce different sources of variability.

Which biospecimen matrices are specifically mentioned?

The NOFO specifically calls out measurement standardization across multiple sample matrices and gives examples including plasma, tissue, and serum extracts.

Why does the NOFO emphasize multiple matrices like plasma, tissue, and serum?

Different matrices can behave differently in measurement workflows (for example, due to matrix effects). Standardizing across matrices helps ensure results are comparable and interpretable when clinical trials use different biospecimen types or when multi-site studies process samples differently.

What does assay calibration and measurement standardization mean in this program?

In this context, it means establishing approaches that allow measurements to be comparable across settings and sites, including addressing matrix effects, using internal standards, applying reference materials, and designing for inter-lab comparability so that results align when the same underlying biological material is tested.

What is meant by "harmonized protocols"?

Harmonized protocols are standardized procedures designed to be used consistently across labs and clinical trial settings. The intent is to avoid each group creating its own incompatible "standard" and instead produce aligned methodologies that the broader clinical research community can use.

Does this program allow animal (preclinical) work?

Yes, but only in a limited and targeted way. Preclinical animal work is allowed only when it is clearly integral to protocol development and specifically needed to test elements of standardized workflows before moving into human testing.

What kind of animal work is considered appropriate under this NOFO?

The NOFO describes targeted work that de-risks or optimizes methodology components, such as evaluating sample stabilization steps, testing extraction approaches in relevant tissues, or assessing how handling variables affect metabolite stability, with the purpose of supporting confident validation in humans.

What kind of animal work is not the intent of this program?

The intent is not to fund broad animal biology studies of NAD. Any preclinical work should be in service of methodology development and validation rather than exploratory or broad mechanistic animal studies.

What is the funding mechanism used for this opportunity?

The funding mechanism is a U01 cooperative agreement.

What does a U01 cooperative agreement imply for how the project will be run?

A U01 cooperative agreement signals substantial scientific and programmatic involvement by NIH compared to a standard investigator-initiated grant. Awardees should expect coordinated engagement consistent with the cooperative nature of the mechanism.

Are awardees expected to work independently or as a group?

Even though projects are funded as individual awards, awardees are expected to function as a coordinated network. The network structure is meant to speed harmonization by requiring active communication, sharing emerging findings, comparing performance across methods and sites, and collectively working through implementation pitfalls.

What is the intended deliverable of the awardee network?

The intent is to produce aligned, validated, shareable, and clinically workable measurement standards for NAD+ and related metabolites that the broader clinical trial community can use, rather than producing multiple incompatible "standards" across different awardees.

Is this opportunity a clinical trial program?

Yes. The title indicates "Clinical Trial Required," and the focus is on methodologies that can be used in human clinical research and clinical trials.

Which agency administers this program?

The program is administered by the National Institutes of Health (NIH).

How is this opportunity categorized in federal assistance terms?

It is described as a discretionary health funding program with CFDA 93.866.

Who is eligible to apply?

The NOFO lists a broad set of eligible applicant organizations, including U.S. governmental entities (state, county, city/township governments, special district governments), public and state-controlled institutions of higher education, private institutions of higher education, independent school districts, public housing authorities/Indian housing authorities, federally recognized Native American tribal governments, tribal organizations (including those not federally recognized), nonprofits with and without 501(c)(3) status, for-profit organizations (other than small businesses), and small businesses.

Does the NOFO call out any specific institution types as eligible?

Yes. It explicitly highlights Alaska Native and Native Hawaiian Serving Institutions, AANAPISI institutions, Hispanic-serving institutions, HBCUs, Tribally Controlled Colleges and Universities, faith-based or community-based organizations, regional organizations, eligible federal agencies, and U.S. territories or possessions.

Are foreign (non-U.S.) organizations eligible to apply as the applicant?

No. Non-domestic (non-U.S.) entities (foreign organizations and foreign institutions) are not eligible to apply as the applicant organization.

Are non-domestic components of U.S. organizations eligible to apply?

No. Non-domestic components of U.S. organizations are also not eligible to apply.

Can a U.S. applicant include foreign components in the project?

Yes, foreign components are allowed when they meet the NIH definition in the NIH Grants Policy Statement. This generally means a U.S. applicant may include well-justified foreign elements as part of the project if they are necessary and appropriately structured, even though a foreign institution cannot be the primary applicant.

What are the key dates mentioned in the provided information?

The provided information lists a creation date of July 5, 2023, and an original closing date of November 2, 2023.

Is the award ceiling provided?

No. The award ceiling was not specified in the excerpted information.

Is the expected number of awards provided?

No. The expected number of awards was not specified in the excerpted information.

What is the broader impact this program is aiming for in NAD+ clinical research?

By producing validated, standardized, and shareable measurement methodologies, the program aims to enable future clinical trials and multi-site studies to interpret NAD-related endpoints with much higher confidence and far less method-driven variability across sites and studies.