Microgravity and bone cells

A CSA-CIHR joint initiative

Requests for Applications

Summary

The Canadian Space Agency (CSA) and Canadian Institutes of Health Research (CIHR) – Institute of Musculoskeletal Health and Arthritis (IMHA) are pleased to announce a Request for Applications (RFA) to researchers with expertise in bone cell biology, particularly in relation to the effects of microgravity on bone loss.

The experiments will take place in the e-OSTEO cell culture "Mini-Lab," which is a self-contained bioreactor system that allows automated growth of cells, and the addition of fluids to the bioreactors from four independent reservoirs. The e-OSTEO system will be described in more detail below.

• Specific objectives and eligible research areas
• Partners
• Eligibility criteria
• Specific eligibility requirements
• Experimental hardware
• Funds available
• Scope and timeline of the proposal
• Budgetary guidelines
• How to apply
• Evaluation process and criteria for peer review
• Deliverables
• Address for submitting application and contact information

Specific objectives and eligible research areas

CSA and CIHR–IMHA are soliciting research that addresses the objectives of the Life Sciences program:

• to use the space environment to better understand life
• to better understand how biological systems and whole organisms, including humans, function and adapt to the environment of space and readapt to conditions on Earth
• to obtain knowledge and develop technology to produce safer space travel and improve life on Earth

This Request for Applications relates to the following question, which is a key component of the CSA's Space Life Sciences program. What are the mechanisms whereby bone loss occurs in space and are these mechanisms similar to those underlying osteoporosis? How can this bone loss be reduced or halted?

Since the Gemini missions in the 1960s, it has been known that bone loss occurs during space flight. In long-term missions, astronauts typically lose one to two per cent of their bone mass per month in space. This bone loss continues for several months after landing, and is of considerable concern because of the increased risk of fractures, particularly in the lower extremities. Also, the loss of calcium from bones imposes an increased load on kidneys, increasing the risk of renal stones and other renal problems.

The cellular mechanism of increased bone loss in microgravity is currently unknown; thus there is considerable incentive to undertake investigations that compare bone cell biology in the absence and presence of gravity.

Since bone loss is an important health concern on Earth, because of the prevalence of osteoporosis, research into the mechanisms of bone loss in space may well increase understanding of bone physiology and disease processes on the ground.

Tissue engineering research is another area of interest for CSA and CIHR. Sometimes, cells grown in microgravity are able to form tissues more closely resembling natural tissues than those formed under the influence of gravity. The e-OSTEO system can accommodate both two-dimensional (e.g., thin films) and three-dimensional growth substrates (e.g., scaffolds), providing an opportunity for tissue engineering research related to bone.

Thus the two main research areas are bone demineralization and bone tissue engineering. All studies must require the environment of space to address or support critical scientific questions. While space flight experiments may include a ground component that complements or supports the flight experiment, studies relying solely on ground experimentation are not within the scope of this announcement. Ground-only studies should be submitted in response to the CSA Announcement of Opportunity for Concept and Feasibility Studies.

Partners

This RFA is a joint initiative of the Canadian Space Agency (CSA) and Canadian Institutes of Health Research (CIHR) - Institute of Musculoskeletal Health and Arthritis (IMHA).

Eligibility criteria

• Proposals will only be accepted from Canadian citizens and landed immigrants. The CSA policy and guidelines for Announcements of Opportunity apply to this competition.
  
• Research teams are encouraged to apply to this initiative.
  
• A concise and complete summary that explains how the proposed flight experiment's objectives can be achieved using the e-OSTEO system should be included as part of the research proposal.

Specific eligibility requirements

The application will cover research leading to and including a flight experiment on a recoverable satellite, which is projected to occur in October 2006, and can also include research involving samples collected from the satellite after landing.

Recoverable satellites are automated systems that are launched by a rocket; they then experience a period of freefall around the Earth. During this time the satellite experiences microgravity. At the end of the freefall period (usually two weeks), the satellite uses small rockets to guide it back through the atmosphere to the Earth. As the satellite is equipped with parachutes, it is usually possible to recover the satellite and its contents with minimal damage.

Because of the unusual nature of the experimental environment, it is important for applicants to be aware of the constraints affecting the flight experiment. The most important guidelines for applicants are:

• Flight experiments should have a strong foundation derived from ground experiments; because of the tight time frame of this flight opportunity, there will be minimal time for ground experimentation before the flight. Therefore, the application should consist of mature studies based on a strong core of current research knowledge.
• The proponents should specify the exact duration of the flight experiment, and should indicate if there is flexibility in this timeframe. The anticipated length of the satellite's orbit around the Earth is 15 days.
• The flight experiment will occur in the e-OSTEO mini-lab for cell culture. Details on the e-OSTEO system are given below, under Experimental hardware. It is unlikely that additional hardware orequipment specific to the proposed research can be developed in the time frame of this flight opportunity.

Experimental hardware

The e-OSTEO system (Figure 1) has four independent trays that will be available to Canadian researchers. Each tray contains three bioreactors, and four syringe reservoirs that can be used to deliver fluid independently to any of the bioreactors. Three of the four syringe reservoirs contain 60 mL volume, and the fourth has 120 mL. Since the reservoirs have a common line feeding the three bioreactors, one of the syringes typically contains a flushing solution, to eliminate the possibility of cross-contamination of the bioreactors. One of the syringes is often used to deliver a chemical fixative to terminate experiments; however, if it is possible to preserve the cells or biochemicals through cooling to 4°C, the use of chemical fixatives can be avoided.

During each experiment, three samples of 3 mL each can be taken from the bioreactors. There is flexibility in the source of each sample. For example, all three samples could be taken from the same bioreactor, or each of the bioreactors could be sampled once. Experimenters have control over the temperature of each tray. For example, if it is desirable to keep cells in a state of low metabolic activity before launch, the tray can be held below 37°C. When the satellite is in orbit, the temperature can then be raised to 37°C. Media samples taken from the bioreactors can be cooled (4°C); however, the bioreactors will also be cooled if this action is taken.

Each bioreactor has a volume of 5 mL, and can hold one standard microscope slide. In past OSTEO experiments, two types of slides have been used. One (2-D) has 16 wells on a slide. Each well has a thin film (5 µm) of Skelite, a synthetic substance that mimics the chemical and physical properties of bone. The other type of slide (3-D) typically has four cylindrical scaffolds (6 mm diameter; 4 mm height) constructed of Skelite. These porous scaffolds have been shown to support bone cell growth. Investigators are not limited to these substrates, as long as the proposed experiment can be accomplished on a microscope slide, and any three-dimensional scaffolds do not exceed 4 mm in height.

Investigators should propose a flight experiment based on one tray (i.e. three bioreactors). If it is vital that two trays are required for the proposed research, the applicant must explain and justify this requirement.

Figure 1. The OSTEO that was used for STS-107 (Columbia).
The system that will be used in the proposed flight
experiment is an upgraded version of this.
(Photo: Millenium Biologix)

The following requirements are specific to the logistics of the flight experiment, and must be addressed in the proposal.

• Experimental materials, including living cells, must be able to survive a period of 106 hours at a temperature of 10 to 37ºC before launch. It is possible to cool the bioreactors and collected samples to 4°C during reentry and recovery phases. Cells and/or collected samples must be able to withstand up to 48 h at this temperature during the satellite recovery phase (i.e. finding and removing the satellite after it has returned to Earth).
  
• There are numerous restrictions on the types of materials that can be used in flight experiments, and in general, chemicals with low toxicity are preferred. When toxic chemicals are essential (e.g. for fixing cells), toxicity evaluations will be done on an individual basis. Please note that specific information on the permissible toxicity of chemicals is currently being collected. This RFA will be updated and applicants notified when this information is available.

Funds available

Financial support is available in the form of contracts of up to $116,667 per annum. The maximum funding available for one proposal is $350,000 for a three-year study (including overhead, benefits, etc., but excluding GST). The number of contracts awarded as a result of this solicitation will depend on the merit of the proposals and the availability of funds.

Scope and timeline of the proposal

Applicants should use the following schedule to base their proposals.

1. Ground testing
   May 1, 2005 to May 1, 2006

   During this phase, successful applicants will receive one e-OSTEO tray for ground testing of their flight experiment. This will allow researchers to acquire expertise with the e-OSTEO software and hardware (researchers will be trained by Millenium Biologix personnel). During this time, researchers can test their experiment, to ensure that the cells and cell products can withstand the conditions (e.g., time and temperature) experienced during the three phases of the flight experiment (pre-launch, orbit, and recovery).

2. Science verification test (SVT)
   May 1, 2006 to Nov., 2006

   This test is a mock-up of the flight experiment. Unlike the ground testing, which is done with individual trays in each researcher's laboratory, the SVT is held in the actual e-OSTEO box that will be used in flight (i.e., all trays will be present).

   Following the SVT, there is typically a small period of time to allow ground experiments to fine-tune the flight experiment.

3. Flight experiment onboard a recoverable satellite
   Target timeframe is during October 2006

   Investigators will fly to a laboratory in Europe, where they will prepare their tray for the flight experiment. The e-OSTEO container will then be flown to the launch site, where it will be loaded into the satellite prior to launch.

   The satellite will experience microgravity for 15 days. It will then return to Earth, and the e-OSTEO container will be recovered and returned to the laboratory in Europe. The investigators will then return to Canada with their cells and/or cell products.

   During the flight experiment, an identical experiment will occur on the ground, using the same e-OSTEO system. This will permit direct comparison of cells experiencing microgravity to cells on Earth.

4. Analysis of experimental outcomes
   End of flight experiment to April 30, 2008

   Investigators can propose follow-up experiments for this time period, or it can be used solely for analysis of the cells and/or cell products of the flight experiment.

Budgetary guidelines

The proposed budget and schedule/milestones for the full duration of the study must be provided.

The cost of equipment required may be included provided it is essential to the project, but it must not exceed $50,000.

All costs of producing anticipated contract deliverables (see below) should be included.

Principal investigators from universities may not include personal salary support in the budget although support for co-investigators (research associates or post-doctoral fellows, not other professors/independent investigators) and other personnel is permitted, and support for students and young researchers is encouraged.

Travel costs to attend conferences should not be included.

The funding of graduate students under this RFA will be permitted only if their supervisors provide written guarantee that they will be provided with the continuous support necessary to complete their graduate studies.

How to apply

• To access CIHR application forms, all applicants and co-applicants must acquire personal identification numbers (PIN). There will be a delay of approximately one working day to receive confirmation of a PIN. More than one working day may be required in peak periods—late January and early August. The PIN will also be needed to validate the Common CV.

• Applicants are advised to refer to the "Guidelines for Completion" specific to each application module, and the instructions on 'How to Apply,' including the applicable Grants and Awards Guides. Note that the proposals must address the criteria described under the "Evaluation Process and Criteria for Peer Review" section in the text of the RFA.

• The following special instructions apply to this RFA:

  To ensure that your registration and application are forwarded to the appropriate CIHR staff, indicate 'Microgravity and Bone Cells: A CSA-CIHR Joint Initiative' under the heading "Name of Strategic Initiative (RFA)" (if applicable) in the General Information section of the Research Module (page 7 of the pdf form).

There are two steps to applying for this RFA. Applicants are encouraged to use the Webforms.

1. Register your intent to apply, using the CIHR Operating Grants registration package, which consists of pages from the CIHR Research Module, in addition to pages one and two of the Common CV for each applicant and co-applicant participating in the research project. Using the Webforms, after entering your PIN and password, create a new application, and then complete the "Operating" section under the Research Funding Programs heading. Once you have completed this section, save and exit, then choose the print option for "Registration Pages Only." Send the original, plus one copy of both the registration pages and the CV pages to CIHR by courier by November 1, 2004.

2. Complete the full application using the CIHR Operating Grants application package, which consists of the CIHR Research Module, the CIHR Operating Budget Module and a Common CV for each applicant and co-applicant participating in the research project. When completing the CIHR Operating Budget Module provide a detailed justification of all project costs. Costs to be covered by CIHR's funding partners should be listed in the "Other Funding Sources" column on page one of the Budget Module. Send the original by courier, including eight copies of the full application by December 15, 2004.

Evaluation process and criteria for peer review

Each proposal must describe how the grant will address one of the important health issues described under "Specific Objectives and Eligible Research Areas" above.

Applications will be evaluated by a CIHR peer-review committee. The committee will be created specifically for this Initiative. Committee members are selected based on suggestions from many sources including the Institute(s) and partner(s). Names of committee members will be published on the CIHR website. The committee will follow the CIHR Peer Review Process.

Following the peer review process, there will be a technical feasibility review led by the CSA. The purpose of this is to ensure the science can be successfully flown within the technical constraints of the flight mission. Further details on the technical review will be provided to the applicants.

Answers to the following questions about the proposed research will be reviewed 

• How important and/or original are the hypotheses or the questions to be addressed, and how clearly are they formulated?
• How important and original are the contributions expected from the research proposed? What is the potential for important new observations or knowledge?
• How well will the proposed experiments address the hypotheses or questions? How appropriate are the methods to be applied and the proposed analyses of data?
• How well will the applicant implement new methods that are to be introduced and/or explored? How well have the applicants anticipated difficulties in their approach and considered alternatives?
• How critically is the relevant literature appraised and evaluated?

The applicant's productivity, experience, and training will be assessed by answering the following questions

• How appropriate to the research proposed is the training or track record of the applicant(s)?
• How important and original is the recent productivity of the applicant(s)?
• How much confidence do you have that the applicant can do the work proposed? (The proposed time commitment to the research may influence this judgment.)

Deliverables

All contracts resulting from this Request for Applications shall include at least the following deliverables:

• three quarterly reports and one annual progress report per year (5 paper copies)
• a final report (5 paper copies and one electronic copy)

In addition, although the following cannot logistically be included as contract deliverables, the Canadian Space Agency requests the receipt of the following, whether they occur during the term of the contract/study, or after its completion:

• copy of scientific papers subsequently published in peer-reviewed journals that may result from work done under this contract.
• description of awards resulting in whole or in part from work conducted under this contract.
• names of persons receiving post-graduate degrees using results in whole or in part conducted under this contract, and type of degree.

Due to the research and development nature of the proposed work, any resultant contract that may result from this RFA will be subject to the most recent General Conditions Research and Development DSS-MAS 9624 (note: Employment Equity only applies to requirements over $200K). Submission of a proposal acknowledges the Bidder's agreement with the application of the DSS-MAS 9624 General Conditions - Research and Development to the contract work. Any resultant contract will be negotiated and issued through Public Works and Government Services Canada (PWGSC).

Any contract resulting from this RFA will contain a clause pertaining to communications/public affairs defining contractor responsibilities and authorities. The CSA retains the right to make primary contract announcements. Any subsequent contract-related announcements will be undertaken in cooperation with and subject to the approval of the CSA.

The basis of payment for the contracts awarded from this RFA will be firm fixed price, milestone payments. Annual milestones, such as at the successful conclusion of the annual review and annual progress report, are preferable.

Proposals must identify any intellectual property that the proposed work is based on and any potential intellectual property that would result from the work.

Address for submitting application and contact information

Send application by courier to:

RE: "Microgravity and Bone Cells: A CSA-CIHR Joint Initiative"
Canadian Institutes of Health Research
Research Portfolio
160 Elgin Street, Room 97
Address Locator 4809A
Ottawa, ON  K1A 0W9

Contacts:

For questions regarding the review process, application forms and submission requirements, please contact:

Michelle Hume
Program Delivery Officer
Canadian Institutes of Health Research
160 Elgin Street, Room 97
Address locator 4809A
Ottawa, ON  K1A 0W9

Tel.: (613) 957-8842
Fax: (613) 954-1800
E-mail: mhume@irsc-cihr.gc.ca

For information about this initiative and CSA, please contact:

Dr. Nicole Buckley
Director, Life and Physical Sciences
Space Science Program
Canadian Space Agency
6767 Route de l'Aéroport
Saint-Hubert, QC  J3Y 8Y9

Tel.: (450) 926-6574
Fax: (450) 926-4766
E-mail: lifesciences@space.gc.ca

For general information about the Institute of Musculoskeletal Health and Arthritis (IMHA), please contact:

Sophia Tsouros
Project Manager/Analyst
Institute of Musculoskeletal Health and Arthritis
160 Elgin Street, 9th Floor
Address Locator 4809A
Ottawa, ON  K1A 0W9

Tel.: (613) 954-3469
Fax: (613) 954-1800
E-mail: stsouros@irsc-cihr.gc.ca