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Contract Topics
274 Quantitative Cell-Based Imaging for Clinical Diagnosis and Treatment
Number of anticipated awards: 3
(Fast-Track proposals will be accepted.)
Budget (total costs): Phase I: $150,000;
Phase II: $1,000,000
(Note: It is strongly suggested that Proposals adhere to the above budget amounts. Proposals with budgets exceeding the above amounts may not be funded. Phase I project periods may last a maximum of 9 months.)
The deadline for receipt of all contract proposals submitted in response to this solicitation is: November 9, 2009.
Summary:
This SBIR announcement solicits proposals to develop semi-automated quantitative imaging assays to enumerate cells in tumor tissue. While genetic, epigenetic or proteomic profiles of tumors may predict therapeutic response, these methods are often slow, complicated and fail to assess small subsets such as cancer stem cells, host immune or stromal cells whose presence may determine the clinical outcome of a treatment. Additionally, this SBIR announcement seeks to standardize objective measurement of immunohistochemistry and in situ hybridization—two processes that have been difficult to standardize. A semi-automated assay also may improve the pathologist's throughput and accuracy compared to manual enumeration of rare cells. These objectives complement several NCI strategic goals in molecular medicine, nanotechnology, and development of novel therapeutics.
SBIR respondents will develop a set of fluorophore-labeled secondary reagents that detect haptens conjugated to primary antibodies or nucleotide probes that react with specific cellular antigens, genes, or gene transcripts. The secondary reagents will be used in standardized tissue-based immunofluorescence assays that can be measured with current state of the art image acquisition and quantitative analysis and that are independent of the antigens or genes being measured so long as the primary reagents bear the appropriate haptens. Preclinical models may be used in Phase I but ultimately the assay must be clinically useful. The multiplex reagents may be provided by a central laboratory in specific assays or distributed through kits. Small businesses may also submit proposals to measure cellular subsets relevant to oncology that they have identified.
Project Goals:
The short-term goal is to develop multiplex reagents to facilitate performance of validated quantitative imaging assays to measure cell subsets within neoplastic tissue. This solicitation seeks to create stable mixtures of fluorescent reagents that detect at least 3 separate and distinct haptens conjugated to primary antibodies or oligonucleotides that are used in immunohistochemistry and/or in situ hybridization. The aim is to create a mixture of three or more secondary reagents that can be used reproducibly in assays independent of the specific antigens or nucleic acids being detected by the primary antibodies or nucleotide probes. The fluorophores on the reagents must be chosen so as to maximize separation of signals by the three or more reagents as well as other reagents that may be used to identify such cell constituents as nuclei. Tissues used for assay development should be collected in accordance with guidelines provided by the NCI Office of Biorepositories and Biospecimen Research (OBBR, please see http://biospecimens.cancer.gov/). The assay can be envisioned as either a service by a central laboratory or as a kit with equipment and supplies, including calibrators and a SOP. SOPs for these assays must be developed and provided to the NCI. Also data using a second technology that confirms the quantitative measurements of the specified cell subsets (e.g., by flow cytometry, functional assay or laser capture microscopy and western blotting) must also be provided to the NCI. Small businesses may submit proposals for the development of assays that have been identified in the literature or by the small business to support development of oncologic therapeutics.
The long-term goal of this contract topic is to develop imaging assays for cell-based markers that provide prognostic, predictive, or therapeutic response information on clinical tissue biopsies that may be used in clinical practice. Achievement of this goal would support the goal of the NCI SBIR program to fund small businesses to develop commercially viable products that advance the research and development needs of the National Cancer Institute. Strategic priorities 4.1-.3 and 4.5 of the NCI Strategic Plan identify efforts to validate biomarkers and targets for cancer prognosis, metastasis, treatment response and cancer progression, then integrating these into clinical trials as being important.
Market analysis indicates that development of quantitative imaging assays is a valuable first step for eventual commercialization of cancer diagnostics and laboratory assays, as well as serving the needs of cancer therapeutic development. In addition to the assay itself, the contract recipient will develop and provide SOPs to the NCI that have been fully qualified or validated with human tumor/tissue samples.
Finally, companies should extend this work into developing either services or kits with integrated equipment that will stratify patients for clinical trial selection or that predict or evaluate response to new therapeutic agents.
- Develop stable mixtures of at least three fluorophore-labeled reagents that stoichiometrically bind to such standard haptens as biotin, dioxygenin, DNP, fluorescein of other compounds conjugated to primary antibodies or nucleotides (please see list below for examples). These reagents should be used in a multiplex quantitative imaging assay that identifies and counts defined cellular subsets in a preclinical model or clinical specimens that is consistent with the current DICOM standard (http://medical.nema.org/Dicom/). The NCI is interested in quantitative multiplex fluorescent assays to enumerate and identify defined cellular subsets in tissue sections of tumors (Please see below for this list of candidates). Each cell type may be identified by separate sets of primary antibodies or oligonucleotides that are individually labeled with one of the haptens that are recognized by the anti-hapten secondary reagents. The aim is to create a mixture of secondary reagents that can be used reproducibly in assays independent of the exact epitopes of the antigens or sequences of the nucleic acids being measured as well as being used in assays that simultaneously measure antigens and genes. The fluorophores on the reagents must be chosen so as to maximize separation of signals by the three reagents as well as other reagents that may be used to identify such cell constituents as nuclei. The imaging assay must demonstrate accuracy in formalin-fixed tissues. The SBIR recipient must deliver SOPs for specimen collection, processing, and stability of the cell subset under the conditions of the imaging assay.
- The SBIR recipient must demonstrate research assay reproducibility, variability, and accuracy in a preclinical model or appropriate tissues for the intended clinical use of the assay. An important aspect of this is the development of reliable calibrators for the assay. The data demonstrating consistency with the current DICOM standard and assay SOPs must be provided to the NCI. The NCI can assist in acquisition of appropriate tissues.
- The SBIR recipient must provide evidence of confirmation of the specificity of the assay's results by a second technology that assesses phenotype or function of the cell of interest.
Phase II activities and expected deliverables:
- Report to NCI confirmation of the use of a stable set of at least three fluorophore-labeled secondary reagents in a semi-automated quantitative imaging assay as described and required in Phase I for a specified cell subset in clinical samples. In addition, a second technology must confirm the measurement of the specified cell subset phenotypically and/or functionally. NCI can assist in acquisition of appropriate specimens for this confirmation and testing. This is expected to require analysis of normal and malignant tissues that are both fresh and formalin-fixed to enable parallel confirmation of phenotype and/or function.
- Provide to NCI the SOP for specimen collection and processing and of the imaging assay for the cell subset with the exception of proprietary information. This SOP should describe the characteristics for stability and availability of all reagents and calibrators.
- Demonstrate acceptable real-time assay performance in clinical tissues appropriate for the intended use of the assay.
- If the product will be a research kit, the kits must be provided to NCI for evaluation. Kits should include:
- Package insert
- Critical reagents
- Calibrator slides
Cell Types and Haptens:
Candidate cell types to be measured by a minimum of three markers
Cancer Stem Cells (ALDH1A1, CD133, CD44v6)
Macrophages
Endothelial Cells
Endothelial Cells with specific receptors (VEGFRs, EGFR, PDGFRs, ALK, etc.)
T cells
B Cells
Memory B Cells
Dendritic Cells
Tregs
TRECs
Other T cell subsets
Other B cell subsets
MDSC
Specific cancer cell subsets (e.g., HER2+ER-, HER2-ER-PR-, Pathway specific Smo, PTCH1, Gli1 etc.)
Candidate haptens as examples to which secondary reagents might be directed
Biotin
Avidin
Dioxygenin
DNP
Fluorescein
Oregon Green
