Regenerative Medicine (RM) using adult stem cells is an emerging area of science that holds great promise for treating and curing a variety of injuries and diseases. While our understanding of adult stem cell biology is progressing rapidly, the need for a more complete understanding of the biological attributes of stem cell products is widely acknowledged, as it may enable the correlation of clinical outcomes with particular cell characteristics, inform future product development, help improve product safety and efficacy, and enhance reproducibility of studies. The purpose of this announcement is to solicit applications from providers of technology platforms and scientific research services to perform high quality in-depth cell characterization (IDCC) assays as outlined below in support of the Regenerative Medicine Innovation Project (RMIP) and to collaborate in the development of approaches for IDCC involving regenerative medicine clinical products.
The 21st Century Cures Act, passed in 2016, includes several provisions, pertinent to RM clinical research, that apply to the National Institutes of Health (NIH), the U.S. Food and Drug Administration (FDA), and the National Institute of Standards and Technology (NIST). One such provision is that NIH, in coordination with the FDA, award grants and contracts for clinical research to further the field of RM using adult stem cells. In response, the Regenerative Medicine Innovation Project (RMIP) was launched to support late-stage pre-clinical and IND/IDE-enabling studies, as well as carefully selected clinical trials that will significantly advance the field of RM. NIH and FDA co-hosted a workshop in December 2017 to explore the state of RM science involving adult stem cells, with a focus on approaches to the development of safe and effective RM products. Through this and other national and international expert forums on RM, NIH identified several critical challenges in the advancement of stem cell research. These challenges include the following: limited understanding of the identity and nature of stem cell products used in clinical applications, difficulties in preparation of cGMP-compliant stem cell products, and need for regulatory support to enable submission of well-developed IND/IDE applications. To address these challenges, the NIH, in consultation with the FDA and NIST, established the Regenerative Medicine Innovation Catalyst (RMIC) to provide critical support services for RM research that include IDCC to describe the molecular and biological attributes of source stem cells and stem cell-derived products. Furthermore, the RMIC will develop ways to optimize the sharing and utilization of IDCC data in order to address gaps in knowledge and further the field of RM. The RMIC, through its collaborative network of entities, is designed to address these aforementioned critical challenges and will:
Importantly, the RMIC will promote transparency and ultimately may contribute to reproducibility and standardization of clinical grade stem cell-derived products.
A key component of the RMIC is the In-depth Cell Characterization (IDCC) Hub, administered by the Progenitor Cell Translational Consortium (PCTC) – University of Maryland, Baltimore. The IDCC Hub is the administrative and coordinating support center responsible for soliciting and managing IDCC services to support RMIP awardees. IDCC is defined as the performance of a suite of assays aimed at describing the structural and biological attributes of single cells or cell populations that characterize their identities and are predictive of their functions. The IDCC assays will be performed in parallel with the RMIP study or post-hoc as feasible and appropriate. Because IDCC assays may identify cellular attributes for which the clinical significance is not currently known, these results are not intended necessarily to inform decisions during the conduct of the study, nor are they intended to factor into the normative oversight requirements and processes for the source study. Final assay results will also be returned to the RMIP study investigators as they become available. In order to accelerate the field and inform oversight of future studies, assay results will be made available to the broader research community via the RMIC one year following the end of RMIP awards (or as appropriate).
Applicants funded under this announcement are required to match these federally awarded dollars (total costs: direct costs and indirect costs including facilities and administrative costs) with at least an equal amount (1:1) of non-Federal contributions, as mandated by the 21st Century Cures Act. Qualifying non-Federal contributions may include state and local funding not originating from Federal funds, private-sector investments, in-kind contributions, and donations from foundations. See 45 CFR 75.306, as well as relevant FAQs, for additional details. Applicants will be required to demonstrate that funds and in-kind contributions are committed or available at the time of, and for the duration of, the award. Applications must identify the source, type, and amount of funds proposed to meet the matching requirement and explain how the value of in-kind contributions was determined. Applications must also include letter(s) of support confirming that the required matching contributions (cash or in-kind contributions such as salary, consultant costs, equipment) are available.
The total of all proposed matching contributions of funds, equipment, and services must at least equal the federal award under this solicitation and contribute to at least 50% of total project costs. Any potential contribution of services should be estimated on a time and material basis, and must include an average labor rate, fringe rate, and overhead rate. Any equipment used as an in-kind contribution must have been purchased with private funds and not obtained with a prior federal award. All funded applicants shall have a government-approved or industry standard accounting system by which actual project costs are tracked and reported pursuant to the Cost Share Standard Operating Process/Procedures. This is an absolute requirement to be sure that cost matching obligations are met.
This Solicitation focuses on the large-scale in-depth cell characterization function of the RMIC for a wide range of cells which includes induced pluripotent stem cells, mesenchymal stem cells, placental cells, bone-marrow derived mononuclear cells, and retinal pigment epithelial cells. Proposals to provide scientific research services for the items listed below will be considered. Strong applications will address each of the following items:
Table 1: List of key cell attributes and suggested assays for In-depth Cell Characterization
Cell Attributes |
Example Assays |
Rationale |
Source Stem cell |
Final Clinical-grade Product |
Safety (Donor eligibility determination, sterility, adventitious agent, pyrogenicity-endotoxin, and mycoplasma testing) |
Donor Eligibility Determination: In accordance with 21 CFR 1271 and FDA guidance using FDA licensed, approved, cleared test kits in a qualified laboratory |
Critical for product safety verification and mitigating risk of infectious disease to patient (Required under regulation for allogeneic cellular products)
|
X |
|
Microbiology / Sterility Testing: USP <71> Membrane Filtration, Bacteriostasis and Fungistasis, Direct Inoculation; Rapid Microbial Detection assays (ATP-based bioluminescence, fluorescent labeling, electrical resistance, nucleic acid probes) of acceptable specificity, sensitivity, and accuracy |
Critical for product safety verification and mitigating risk of infectious disease to patient (Required under regulation for cellular products) |
X |
X |
|
Pyrogenicity / Endotoxin Testing: USP <85> Limulus Amebocyte Lysate (LAL) Assay (Kinetic turbidimetric, Chromogenic , Gel-clot) |
X |
X |
||
Virology/Adventitious Agents (Master Cell Bank, Working Cell Bank, End-of-Production): ICH Q5A Viral Safety Evaluation, Retrovirus (cell culture infectivity assays / electron microscopy); in vitro-based assays using susceptible indicator cell cultures (range of human and relevant animal viruses); in vivo, animal-based inoculation assays (viruses not able to grow in cell culture); species-specific antibody production testing; PCR-based direct-testing of cells |
X |
X |
||
Mycoplasma Testing: USP <63> culture-based detection; PCR-based testing (demonstrate comparability to compendial culture-based method)
|
X |
X |
||
Identity (Species determination) and Sex (Male/Female) |
PCR-based assays, Flow cytometry
|
Species determination used as an additional check to confirm authenticity
Sex determination provides for confirmation of authenticity, may be useful for purposes of identification post-administration to assess distribution and possible cell migration |
X |
|
Purity and Heterogeneity |
Single-cell RNA-seq, Single cell ATAC-seq, Flow cytometry, Mass cytometry
|
Used to evaluate the composition profile of cellular preparations and to identify presence of non-target cell types that may pose risks to subjects receiving a cellular product |
X |
X |
Viability |
Trypan Blue Dye exclusion assay, Acridine Orange and Propidium Iodide (AOPI) staining, MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) tetrazolium reduction assay, Electrical potential and Morphometric assays, apoptosis assays, mitochondrial membrane potential-dyes, luminescence ATP assay, oxygen consumption and glycolysis activity |
Critical for determining safety and efficacy of cellular products intended for patient administration (Required under regulation for cellular products) |
X |
X |
Senescence |
Telomere length assay
|
For limited replication capacity cell phenotypes with functional proper-ties that degrade with increasing passage number as proliferation limits are reached |
X |
X |
Genetic Stability |
Karyotyping, Southern blot analysis, FISH assays
|
For cellular phenotypes exhibiting a high proliferative capacity that may accumulate genetic mutations over extended periods in culture that impact functional properties or pose a safety risk if given to patients |
X |
X* |
Cell Proliferation |
MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) tetrazolium reduction assay, Alamar Blue, detection of cellular proliferation markers (Ki-67, PCNA [proliferating cell nuclear antigen], topoisomerase IIB), luminescence ATP assay |
Serve to identify cellular products that could pose a safety risk to subjects. Typically performed in conjunction with preclinical toxicology testing |
|
X |
Clonal Capacity
|
In vitro single-cell clonogenic assays |
Determine ability of stem cell self-renewal and generation of differentiating lineages |
X |
|
Pluripotency/ Multipotency
|
Quantitative RT-PCR assessment of Oct4, Sox2, Lin28, Klf4, and c-Myc genes among others; Flow cytometric assessment of Oct4, Lin28, SSEA4, TRA-1-81, TRA-1-60, Sox17 proteins among others
Quantitative assessment of differentiation capacity; Embryoid body formation, embryonic lineage commitment, specific lineage differentiation (i.e., cardiac, neuronal etc.); Retinal pigment epithelium commitment; Adipogenesis, chondrogenesis, or osteogenesis for MSCs |
Provide useful information concerning a key attribute of starting cellular material for the purpose of establishing whether there is a safety risk that may be posed by the presence of residual, non-target pluripotent stem cells in the final clinical-grade product
Provide quantitative assessment of specific function
|
X |
X |
Tumorigenicity
|
OncoPanel assay and FDA-approved F1CDx and MSK-IMPACT Next Generation Sequencing (NGS) to detect tumor gene alterations (MSK-IMPACT is a single-site assay performed at Memorial Sloan Kettering Cancer Center) |
Provide information concerning the potential for a final clinical-grade cellular product to exhibit neoplasia following administration to subjects. Confirm results through preclinical toxicology testing |
X |
X |
Transcriptome
|
Single-cell RNA-seq, Single molecule-FISH |
Use information to correlate with quantitative biological function and clinical outcome assessments |
X |
X |
Epigenome
|
ATAC-seq, ChIP-seq, Methylome assays |
Use information to correlate with quantitative biological function and clinical outcome assessments |
X |
X |
Genome
|
Next Generation Sequencing (NGS), including assays to detect SNP and SNV |
Assess or compare genetic stability |
X |
X |
Proteome
|
Liquid Chromatography Mass Spectrometry (LC-MS), Nuclear Magnetic Resonance (NMR) spectroscopy, Gas Chromatography Mass Spectrometry (GC-MS), Mass cytometry, CyTOF |
Use information to correlate with quantitative biological function and clinical outcome assessments |
|
X |
Lipidome and Metabolome
|
Mass spectrometry and Mass cytometry-based assays |
Use information to correlate with quantitative biological function and clinical outcome assessments |
|
X |
Comprehensive Assessment of Cellular Markers (intracellular and cell surface). |
Flow cytometry (Including but not limited to assessment of markers that are critical for cell survival and engraftment, immune suppression and evasion, and differentiation), Electron microscopy (SEM/TEM) to characterize apical/basolateral morphology
|
Useful for the stereotypic identification of cellular phenotype(s) based on assessment of select markers associated with critical functional attributes indicative of biologic activity or indices of risk posed to subject safety. Also, can correlate with other transcriptome and possibly other omics analysis |
X |
X |
Potency
|
Examples of cell-specific functional assays include cytokine secretion profile analysis, flow cytometry for canonical cell markers, cell polarization, electrical resistance and stereotypic morphometric analysis |
Critical for determining efficacy of cellular products intended for patient administration (Required under regulation for cellular products) |
|
X |
* One-time testing of final clinical-grade cellular products requiring additional cell expansion from each Working Cell Bank
Furthermore, applicants must address the following criteria for assuring quality and comparability of generated assay data:
Applicants must be willing to confer with the National Institute of Standards and Technologies (NIST) to identify measurement science and measurement assurance strategies for each of the measurement platforms so that appropriateness of the assay method and confidence in the assay result can be determined. Please see https://www.nist.gov/mml/bbd/cell-measurements/advanced-therapies-cellular-and-gene-therapy-and-regenerative-medicine-2 for more details on NIST’s biosystems and biomaterials measurement assurance programs.
To ensure that data can be shared broadly across databases and optimally used across research, it is important for applicants to utilize appropriate data standards and common data elements to the extent possible which would help to maximize the value and interoperability of data in advancing research. Accordingly, when developing and collecting data and data sets, awardees should consider following the NIH resource guide document which provides sample tools and information for identifying and utilizing certain data standards and common data elements in NIH programs. Furthermore, awardees are expected to adhere to data standards that are developed in collaboration between IDCC Hub, RMIC, NIH, FDA, and NIST.
Applicants must accept that ownership of the provided data is retained by the originating entity. Service providers will not use, release, reproduce, distribute, or publish any of the data produced in the conduct of IDCC assays, nor authorize others to do so, without written permission of the NIH or other named entities. The awardees will be expected to follow all applicable NIH regulations, policies, and expectations concerning sharing research data, biomedical materials, and publication, as well as any specific requirements within the RMIP funding opportunity announcements and the terms of individual awards. Furthermore, awardees are expected to follow NIH standards for overall data quality, transfer, and management.
The purpose of the submission is to provide the IDCC Hub with information on proposed capabilities and scope of services. A solicitation timeline is included below. Instructions on what to include in the application, and the technical evaluation criteria by which submissions will be assessed are found in the Submission Evaluation Criteria section.
Prospective applicants are asked but not required to submit a letter of intent that includes the following information:
Although a letter of intent is not required, is not binding, and does not enter into the review of a subsequent application, the information that it contains allows IDCCH staff to estimate the potential review workload and plan the review.
The letter of intent is to be sent by August 15, 2020, as a pdf attached to an email to mterrin@som.umaryland.edu with cc to alefever@som.umaryland.edu and litang@som.umaryland.edu
Activity |
Proposed Dates |
Solicitation Announced: |
July 13, 2020 |
Letter of Intent Due: |
August 15, 2020 |
Application Submission Due: |
September 14, 2020 |
Review of Applications: |
November 26, 2020 |
Estimated Award Date: |
December 15, 2020 (earliest possible) |
Use of the submission template will facilitate technical evaluation. The submission will capture the information outlined below. Please use the Solicitation Submission Template outline below to complete your submission:
Applications will be accepted electronically up to 5:00 PM Eastern time on the due date specified. An acknowledgement email will be sent immediately after receipt. Questions and completed Intent to Submit and completed Proposals should be submitted to mterrin@som.umaryland.edu with copy to alefever@som.umaryland.edu and litang@som.umaryland.edu. Proposals that do not follow the submission TEMPLATE will not be accepted.
Submissions will be evaluated using NIH review criteria – Significance, Investigators, Innovation, Approach and Environment -- and prioritized by scientific and technical merit for each of the cell attribute characterization assays offered in the Approach; facilities that include a central facility for performance of characterizations, dedicated space for staff and equipment, and an offsite facility for backup of data; resources to ensure secure internet access; track record and qualifications of the scientific leadership, i.e., principal investigator(s) in performance of in-depth cell characterizations; electronic specimen tracking system; quality management plan; proposed site organization and time lines; and, cost.
Applicants should not include confidential or proprietary information in their applications that they do not want shared within the IDCC Hub. If there is a need to include confidential or proprietary information, please contact us at mterrin@som.umaryland.edu (cc: aminul.islam@nih.gov) to schedule a phone call to discuss further. The IDCC Hub recognizes that commercialization is important to developing products that will ultimately be made available to the public, and thus any proprietary information provided by the applicants or RMIP awardees will be protected to the extent possible under the law.
Communication and questions during the solicitation period (including both submission and evaluation steps) should be directed by email to Dr. Michael Terrin (mterrin@som.umaryland.edu) with copy to Dr. Aminul Islam (aminul.islam@nih.gov) . If the communication/question pertains to a specific proposal under review please provide the proposal title and name of the applicant.
Copyright ©2016 NHLBI Progenitor Cell Translational Consortium.