Q. What are the ages of two group of animals?
A: Forty (40) female BALB/c mice will be launched in two (2) transporters and transferred to four (4) Rodent Habitats on board the ISS and housed for up to ~60 days. Depending upon launch scrub scenarios for SpX CRS-16 that may delay the target launch date of November 27, 2019, twenty (20) mice will be 10-16 weeks old (young) and twenty (20) will be 30-52 weeks old (old) at launch.
Q: Which strain – which ages how many mice. Do you already have some investigators for this mission? are all tissues currently available? Who will judge proposals and determine how the tissues will be distributed? Who will do the dissections? How will the Dissection order be determined.
A: Female BALB/cAnNTac mice from Taconic Biosciences will be the model used for Rodent Research Reference Mission 1 on RR-8.
No investigators or teams or institutions have been pre-selected to receive biospecimens from the mission. Any and all tissues are available for request by respondents to the RFP.
The proposed study has been reviewed and approved internally at the ISS National Lab. As a rodent research reference mission designed to provide flight biospecimens and data to investigators using the Mus musculus BALB/cAnNTac animal model on the ISS National Lab, all biospecimen requests to utilize data or tissues generated from this investigation will be peer-reviewed as part of the the ISS National Lab RFP (Request For Proposals) solicitation process. All proposals received from U.S. based investigators in response to the RFP will be peer-reviewed for technical merit and other selection criteria. This process is described in detail in the ISS National Lab Proposal Submission Instructions and Evaluation Overview (Document dated July 14, 2017, See http://www.spacestationresearch.com/research-on-station/proposal-process/).
All dissections of LAR mice returned to Earth from ISS and ground control counterparts will be performed in a facility or facilities in the southern California area, although the exact location of the facility is still being assessed. Dissection and preservation methods will be determined based on optimizing tissue and data collection outcomes with the specific requirements and needs of the selected proposals in mind. Carcass dissections will be performed at the University of Colorado, Boulder using BioServe labs and facilities.
The order of organ dissection and dissection flow will be determined by the ISS National Lab in accordance with experience from previous RR missions and guidance from subject matter experts at BioServe Space Technologies. Respondents to the RFP are encouraged to include detailed technical information and to cite protocols in their proposal submission for dissection and tissue preservation procedures that optimize tissue and data collection outcomes for assays and technical analyses defined in the proposal for requested biospecimens.
Q: Will they be using the enrichment hut?
A: Yes. The Flight Institutional Animal Care and Use Committee (FIACUC) requires use of the huts as enrichment for the animals.
Q: Will there be a bias in the proposal evaluations? ie academic vs commercial vs government?
A: No. There is no preference or weighting given to the type of authorized institution submitting the proposal.
Q: This RFP is attractive but there is no funding provided for research. As a faculty working in an academic institution, that is very problematic. Is this RFP designed with the biotech industry in mind and not academic institutions? Any suggestions where to get the research funding?
A: The RFP is open to any United States Person as defined in Section 1.4 of the RFP and any United States Person working as part of a US-based laboratory, investigator team, institution or consortium of institutions may apply. Any eligible institutions and/or individuals whose research program can scientifically benefit from receipt of the flight and ground biospecimens and whose proposal defines research objectives that are able to meet the research and technology development objectives under Section 1.5 of the RFP to generate and share data for access by the research community will be considered. The RFP was conceived to be executed to maximize science return from the reference mission to benefit the greatest cross section of research investigators and was not intended to target any one sector of the R&D community. Multi-investigator and multi-institution proposals from self-identified teams of individuals and proposals from research consortia are encouraged.
Q: Can feces samples be preserved in mRNA protecting agents such as RNAprotect or Trizol?
A: Fresh feces can be collected from the LAR (live animal return) mice and their matched ground controls on the ground, and frozen feces can be recovered from the colons of cryopreserved carcasses after their return to Earth.
Q: Will feces be collected on this experiment?
A: Fresh feces can be collected from the LAR (live animal return) mice and their matched ground controls, and frozen feces can be recovered from the colons of carcasses cryopreserved in orbit
Q: What is the ISS National Lab specifically looking for proposals?
A: This Request for Proposals (RFP) solicits applications from investigators from any U.S. based research institution (academic, government, for-profit, not-for-profit) for access to biological specimens (biospecimens) from a Rodent Research Reference Mission to support fundamental, applied and commercial biomedical research on the ISS U.S. National Laboratory.
Q: Can blood draws occur throughout the time that the mice will be on the ISS?
A: No. The baseline Experiment Requirements Document for the mission does not include serial blood collection from the mice during flight operations on the ISS. Blood will be collected in orbit after administration of anesthesia and euthanasia by exsanguination through cardiac puncture followed by thoracotomy. Blood will be separated via centrifugation into serum and cellular components and cryopreserved in orbit for return to Earth.
Q: Will you consider partial funding of some of the specific aims, if they are scientifically distinct, or should we submit separate proposals?
A: No, the ISS National Lab funding is available for this solicitation.
Q: Is it possible to obtain non-fixed spleen samples to potentially extract live immune cells for functional analysis?
A: Yes. It is possible to obtain non-fixed, non-frozen spleen samples from LAR (live animal return) mice. Requirements for additional procedures necessary to maintain biospecimen viability can be included in the proposal for review.
For mice processed in orbit, the spleen will be removed and placed in RNAlater following cardiac puncture and blood draw. The spleen in RNAlater shall be stored for a period of 2 to 14 days at 2-8oC before being frozen at -80 oC or colder until returned to the PI.
Q: For the live animal return, will cell culture be a capability at Long Beach facility?
A: No. The baseline Experiment Requirements Document for the mission does not include cell culture capability at the Long Beach facility for LAR mice. However, requirements for additional procedures necessary to maintain biospecimen viability can be included in the proposal for review.
Q: Can cryo-preserved bone marrow be made available?
A: Bone marrow may be requested, but as the procedures to harvest marrow from bones typically requires destroying both the humerus and the femur to obtain sufficient marrow for analysis, requests for marrow will be weighed against the scientific priority of bones requested for other analyses.
Q: Is it possible to collect urine from the animals aboard the ISS/ground?
A: No. It is not possible to collect urine from the mice on board the ISS. Urine may be collected from LAR mice. Requirements for additional procedures necessary to maintain biospecimen viability from LAR mice can be included in the proposal for review.
Q: At what institution will LAR animals be processed, and will there be capacity for live tissue collection and testing by investigators whose proposals are selected?
A: All dissections of LAR mice returned to Earth from ISS and ground control counterparts will be performed in a facility or facilities in the southern California area, although the exact location of the facility is still being assessed. Dissection and preservation methods will be determined based on optimizing tissue and data collection outcomes with the specific requirements and needs of the selected proposals in mind. Carcass dissections will be performed at the University of Colorado, Boulder using BioServe labs and facilities. Respondents to the RFP are encouraged to include detailed technical information and to cite protocols in their proposal submission for dissection and tissue preservation procedures that optimize tissue and data collection outcomes for assays and technical analyses defined in the proposal for requested biospecimens. Requirements for specific protocols and/or procedures necessary to maintain biospecimen viability and/or sample quality for live tissue collection to support downstream analytical procedures for LAR mice should be included in the proposal for review.
Q: Will intestines and lymph nodes (mesenteric or cervical) be part of the “remaining carcass at -80C” ? In general, where could we find the info on what exactly will be taken from/left into that carcass?
A: Yes. Intestines and lymph nodes will remain in the carcass.
Q: Is it possible to obtain serum from the blood draws?
A: Blood will be separated via centrifugation into serum and cellular components and cryopreserved in orbit for return to Earth. Both layers can be removed after thawing and analyzed.
Q: Does this solicitation require a full ~10 page proposal with a budget even though no funds are being awarded? Thanks!
A: Responses to the RFP do not require the submission of a full 10-page proposal, but responses must include sufficient technical detail and explanation of significant science aims to justify receipt of the biospecimens in response to the research and technology development objectives in Emphasis Area 1 or Emphasis Area 2. Respondents are encouraged to include budget information for the proposed analyses of requested biospecimens.
Q: How many individual samples from the different mice groups can a researcher obtain (to enable statistical interpretation of analytical results)?
A: Respondents may request as many individual biospecimen samples from the mission baseline, ground, and flight groups of mice as required to accept or reject their hypothesis. Power analysis may be included to support the minimum sample size of biospecimens required for a reasonable likelihood to detect an effect of a given magnitude.
Q: Can feces be collected from the animals on the ISS/ground?
A: Fresh feces can be collected from the LAR (live animal return) mice and their matched ground controls, and frozen feces can be recovered from the colons of carcasses cryopreserved in orbit.
Q: So are the requested proposals essentially to describe tissues you would like access to from these mice on RR-8 and the scientific question you are addressing or will the proposal require a more technical assessment of how mice should be processed etc.?
A: Yes. Responses must include sufficient technical detail and explanation of significant science aims to justify receipt of the biospecimens in response to the research and technology development objectives in Emphasis Area 1 or Emphasis Area 2. Technical requirements for additional procedures necessary to maintain biospecimen viability during dissection should be included in the proposal for review.
Q: Is there any possibility of obtaining live mice? My proposal involves scRNA-seq of 20+ organs, but frozen organs cannot be used. We have a team of organ experts ready to dissect and process each organ, and we have an established protocol to get viable cells from all organs simultaneously.
A: Yes. Technical requirements for additional procedures necessary to maintain biospecimen viability during dissection should be included in the proposal for review.
Q: You mentioned that blood will be spun down on the ISS – does this include isolation of plasma and/or PBMCs?
A: Blood will be separated via centrifugation into serum and cellular components and cryopreserved in orbit for return to Earth. Both layers can be removed after thawing and analyzed.
Q: Will the specimens stored in PFA be kept by the ISS National Lab team, or will they be available to be used in proposals?
A: All biospecimens will be available for sharing.
Q: Where will ground control specimens be housed, and will selected investigators have access to these specimens at the time of processing for live tissue functional analysis?
A: Ground control and baseline animals will be housed in the Animal Care Facility at the NASA Kennedy Space Center in Florida. Biospecimen samples from the mission baseline, ground, and flight groups of mice will be available to investigators.
Q: How soon after return to Earth will the LAR mice be sacrificed?
A: LAR mice will be returned to Long Beach, CA by boat and then transported to a select facility by van within 40-55 hours of splashdown. Then, mice will undergo DXA scanning and possibly other physiological and behavioral testing before being processed within 8-24 hours of receipt. The exact timing will depend on specific requirements for tissue, organ, sample and data collection identified in proposals selected for receipt of biospecimens. Every effort will be made to prioritize requirements in such a way as to optimize research outcomes for the investigators.
Q: This RFP does not provide any funding however the ISS National Lab proposal guidelines (section 2.4) refers to a budget. This is confusing. Could you clarify please?
A: The ISS National Lab is not providing any funding in response to this solicitation. Respondents are, however, encouraged to include budget information for the proposed analyses of requested biospecimens, which serves to show the ISS National Lab’s evaluators that the project is funded to such an extent that successful completion of the experiment is reasonably assured.
Q: I received an email saying the webinar started at 3 EST; is that not correct? Nothing seems to be happening…
A: The webinar recording and presentation will be posted online at https://www.issnationallab.org/research-on-the-iss/solicitations/2018-rodent-research/ .
Q: Can experiments be performed on the LAR mice after they arrive in Long Beach or are they exclusively designated for tissue harvesting at that point?
A: Minimally intrusive experiments that may include additional physiological or behavioral testing of the LAR mice may be included in submitted proposals but any such tests will need to be prioritized within the overall timing requirements of other investigators seeking tissues.
Q: Is there a mission 2 planned? If so, would the methods be similar? What would the anticipated date be?
A: There may be additional Rodent Research Reference missions to the ISS National Lab in the future. Please watch https://www.issnationallab.org/.
Q: I am a US citizen, I completed my B.S. and M.S. at Oregon State University under Dr. Russell Turner, whom I collaborated with analyzing rodent spaceflight bone samples from STS-41 as well as my master thesis research. Now, I am conducting my PhD research at SCK•CEN in Belgium in spaceflight immunology.
My question is whether the restriction is related to me being a US citizen or if the research has to be completed at a US-based research institution? If the latter is the case, we have discussed potentially collaborating with my mentors at Oregon State University. As we are conducting a large in-vivo study using rodents to simulate simultaneous spaceflight stressors (microgravity, radiation + psychological stress). Acquiring spaceflight samples, or potential access to NASA life sciences data archive for comparison to the experiments conducted at SCK•CEN.
Would you be able to comment on this question?
A: The RFP is open to any United States Person as defined in Section 1.4 of the RFP and any United States Person working as part of a US-based laboratory, investigator team, institution or consortium of institutions may apply.
Q: I am interested in applying for the Rodent Research Reference Mission-1. I understand from the proposal instructions, that the ISS National Lab will not consider proposals with any non-U.S. entity. As a US citizen based at a non-US institution, I am curious to know if this restriction applies to my proposals. If so, would the addition of a US affiliation to myself or the addition os a US citizen-US based PI to the proposal, change this restriction in any way?
A: The RFP is open to any United States Person as defined in Section 1.4 of the RFP working at a US-based laboratory or institution. In order to execute an unfunded user agreement, the Principal Investigator for the proposal must be a United States Person employed at a US-based institution as certified by an authorized official of the institution.
Q: What is the purpose of having half the mice returned to Earth alive and exposed to Earth’s gravity for up to 48 hours before being euthanized?
A: Access to LAR mice enables investigators to identify in their proposal specific dissection protocols and/or protocols for sample preservation of tissues dissected on Earth that support full processing and preservation of tissues for downstream analyses. In orbit dissection and tissue preservation protocols are more constrained than those available on Earth. The carcass of animals processed in orbit will be frozen at –80oC.
Q: Will investigators have access to intact whole limbs? Or will limbs be dissected and bones and muscles separated before transfer to investigators?
A: Technical requirements for additional procedures necessary to maintain biospecimen viability during dissection can be included in the proposal for review.
Q: What will happen to limbs stored in formaldehyde? Will those samples be available to investigators?
A: These limbs will be returned to Earth and distributed to investigators.
Q: Can investigators ask to have samples stored in specific reagents (e.g. TRIzol)?
A: Technical requirements for additional procedures necessary to maintain biospecimen viability during dissection procedures for LAR mice on the ground can be included in the proposal for review. The use of specific reagents for tissue preservation in orbit, e.g. TRIzol, that are not listed in the baseline Experiment Requirements Document is not possible.
Q: Will DXA scan data be made available to investigators?
A: All mission data will be distributed to the investigators at the earliest possible date and to the entire community within 6-12 months after launch.
Q: Will the ISS National Lab use any of the material or can all tissue go to one investigator?
A: The ISS National Lab will not use any of the material, all tissues will be distributed.
Q: Can one investigator get all limbs?
A: Any and all tissues are available for request by respondents to the RFP.
Q: The webinar was very informative. My research focus is on the epigenetics (e.g., DNA methylation, ncRNAs) of tissue especially brain aging, in healthy and/or stress-related conditions. The impact of microgravity on mouse tissue aging is extremely intriguing for the field to understand the plasticity/resilience of mammalians. We study the aging in all specific brain regions/areas such as frontal cortex, hippocampus, striatum, cerebellum and so on. I thus wonder if the subregions of the brain instead of the whole brain would be dissected for the purpose or research?
A: Any and all tissues are available for request by respondents to the RFP. The RFP is open to any United States Person as defined in Section 1.4 of the RFP and any United States Person working as part of a US-based laboratory, investigator team, institution or consortium of institutions may apply.
Q: Are NASA scientists or civil servants from other government agencies prohibited from submitting proposals and receiving tissues from this mission?
A: NASA scientists and civil servants from other government agencies are allowed and encouraged to submit proposals requesting biospecimens.
Q: In the ISS National Lab budget template what information should be included in other funding/non-ISS National Lab funding section?
A: The other funding/non-funding section should include any costs that your organization and any organization identified as co-investigator or sub-contractor in the proposal will incur as a result of this project.
