Integrating Biospecimen Science Approaches into Clinical Assay Development (U01 Clinical Trial Not Allowed) | PAR 25 325

Frequently Asked Questions (FAQs)

What is the main purpose of this funding opportunity?

This NCI opportunity funds studies that address preanalytical variability in biospecimens. The goal is to understand how specimen collection, handling, processing, transport, and storage affect biomarker measurements, and to develop practical, evidence-based ways to reduce those sources of variation so clinical assay development and analytical validation can be more reliable and efficient.

What problem is NCI trying to solve with this program?

The program targets a common reason clinical biomarker assays fail to perform consistently: differences in what happens to a sample before it ever reaches the assay. Even small changes in preanalytical conditions can change measurable biomarker levels, integrity, or detectability and cause inconsistent results across sites or studies.

What does "preanalytical variability" mean in this context?

Preanalytical variability refers to changes introduced before the analytical testing step, such as differences in how specimens are collected, the time between collection and processing, fixation or stabilization delays, tube types and anticoagulants, centrifugation conditions, temperature exposure during transport, freeze-thaw cycles, storage duration, and storage temperature.

What is the scientific focus of the research expected under this NOFO?

The focus is experimental and mechanistic work that systematically characterizes how specific preanalytical variables affect biomarker readouts across clinically relevant testing platforms, and then uses that evidence to reduce or control those variables in real-world workflows.

Is this opportunity limited to one biospecimen type?

No. The NOFO explicitly supports multiple biospecimen types, including tumor tissue biopsies, blood-based specimens used for liquid biopsy applications, and a wide range of other bodily fluids and specimen types.

What tumor tissue specimens are emphasized?

The NOFO emphasizes clinically common small tumor tissue specimens such as core biopsies and small excision samples, where limited material and rapid degradation can create major challenges for downstream assay performance.

Does this program support liquid biopsy-related biospecimens?

Yes. It supports blood-based biospecimens used for liquid biopsy applications, including studies of how handling and processing variables influence downstream measurement of liquid-biopsy analytes.

What analytes or biomarker classes are mentioned for blood-based specimens?

The opportunity mentions analytes such as circulating tumor DNA, RNA, proteins, extracellular vesicles, metabolites, and immune markers.

What preanalytical factors are highlighted for blood-based biospecimens?

Examples include tube type, time-to-processing, centrifugation conditions, temperature excursions, freeze-thaw cycles, and storage duration, all of which can meaningfully affect measured biomarker signals.

Are non-blood, non-tissue specimen types allowed?

Yes. The NOFO invites studies involving tissue swabs, tissue secretions, pleural and esophageal aspirates, feces, and multiple bodily fluids, reflecting interest in less invasive sampling and multi-omic biomarker development.

Which bodily fluids are explicitly listed as in-scope?

The text lists sweat, urine, cerebrospinal fluid, breast milk, and saliva as examples of bodily fluids of interest, among others.

What kinds of study designs are expected?

Applicants are expected to run controlled comparisons that isolate key preanalytical variables (for example, delaying fixation vs. immediate fixation) and quantify the impact on biomarker readouts using clinically relevant analytical methods.

Is descriptive observational work sufficient for this program?

The opportunity emphasizes experimental and mechanistic approaches rather than purely descriptive work, meaning the intent is to test and quantify cause-and-effect relationships between specific preanalytical conditions and specific assay readouts or failures.

What are examples of research questions that fit this NOFO?

Examples mentioned include how delays in fixation or stabilization influence nucleic acid fragmentation, how different anticoagulants alter downstream measurements, how limited specimen volume affects extraction efficiency, and how storage temperatures and durations change protein stability or microbial signatures.

What deliverables is NCI looking for?

The intended deliverables include practical outputs that can be adopted by laboratories and clinical sites, such as standardized operating procedures (SOPs), acceptance criteria, and best-practice recommendations grounded in evidence linking preanalytical variables to measurement performance.

How does this program aim to improve clinical assay development?

By reducing avoidable preanalytical noise, the program aims to make assay results more reproducible across sites, studies, and patient populations, which in turn can speed the path from biomarker discovery to reliable clinical testing and reduce failures during assay development and analytical validation.

Why is this especially important in oncology biomarker work?

Modern oncology biomarker work often relies on small biopsies, longitudinal sampling, and multi-site studies. In these settings, inconsistent collection and processing can magnify variability and reduce reproducibility, making harmonized preanalytical practices particularly important.

What is the funding mechanism for this opportunity?

The award is a U01 cooperative agreement.

What does a U01 cooperative agreement imply for awardees?

A cooperative agreement generally involves substantial scientific involvement from NIH/NCI staff compared with a standard investigator-initiated research grant, so recipients should expect active engagement from program staff as part of the project.

What does "Clinical Trial Not Allowed" mean here?

It generally means the funded activities should not include prospective assignment of human participants to interventions to evaluate health outcomes. The research may still involve human specimens and associated data, but it should remain focused on biospecimen science, preanalytical variables, and assay-related performance rather than interventional clinical testing.

Can projects use human specimens under this NOFO?

Yes. The description indicates the research can involve human specimens and associated data, as long as the work remains within the non-interventional scope focused on preanalytical conditions and assay performance.

Who is the sponsoring agency and institute?

The agency is the National Institutes of Health (NIH), and the sponsoring institute is the National Cancer Institute (NCI).

What is the opportunity number for this NOFO?

The opportunity number is PAR-25-325.

What is the Assistance Listing (CFDA) number associated with this opportunity?

The Assistance Listing number associated with the opportunity is 93.393.

When is the closing date listed for this opportunity?

The listing shows an original closing date of September 10, 2027.

Is the award ceiling provided in the information given?

No. The award ceiling is not specified in the provided source information.

Is the expected number of awards provided in the information given?

No. The expected number of awards is not specified in the provided source information.

What types of organizations are eligible to apply?

Eligibility is broad and includes domestic organizations such as state and local governments, public and private institutions of higher education, nonprofits (with or without 501(c)(3) status), for-profit organizations (including small businesses), independent school districts, eligible tribal entities, and housing authorities.

Are mission-specific and historically underserved institutions included as eligible applicants?

Yes. The NOFO highlights categories such as HBCUs, Hispanic-serving institutions, tribally controlled colleges and universities, Alaska Native and Native Hawaiian serving institutions, and AANAPISI institutions, among others.

Are faith-based or community-based organizations eligible?

Yes. Faith-based and community-based organizations are listed among eligible applicant types.

Are foreign (non-U.S.) organizations eligible?

Yes. Foreign organizations and regional organizations are listed as eligible.

Can U.S. territories or possessions apply?

Yes. U.S. territories or possessions are indicated as eligible to apply.

Is the program intended for teams with specific expertise?

Yes. The opportunity is aimed at teams that can bridge biospecimen science and assay development, meaning groups that understand clinical realities of specimen collection as well as analytical realities of biomarker measurement.

What makes a project a strong fit based on the description?

Projects are likely to fit well when they identify high-impact real-world preanalytical problems, test them rigorously with controlled comparisons, quantify effects on clinically relevant assay readouts, and produce actionable guidance that can be adopted across laboratories and clinical sites.

Does the NOFO specify which testing platforms must be used?

No specific platform is mandated in the provided description. The focus is on clinically relevant analytical methods and understanding how preanalytical conditions impact biomarker measurements across different testing platforms.

What is the bigger outcome NCI is aiming for?

NCI aims to speed translation from biomarker discovery to reliable clinical testing by reducing avoidable preanalytical noise, improving reproducibility across sites and studies, and supporting harmonized specimen handling practices that reduce measurement failures or biases.