Army 17. 1 Small Business Innovation Research (sbir) Proposal Submission Instructions

PHASE III DUAL USE APPLICATIONS: If a Phase III option is available; the CASEVAC mission constraint system should be targeting toward integrating with current and future medical resupply and CASEVAC UAV/UGV platforms. Platforms for consideration are the U.S. Army Maneuver Center of Excellence (MCoE) autonomous Squad Maneuver Equipment Transport (SMET), The Office of Naval Research (ONR) Autonomous Aerial Cargo/Utility System (AACUS), and Defense Advanced Research Projects Agency (DARPA) Aerial Reconfigurable Embedded System (ARES) to name a few. Reports and documents on the research study and the developed prototype may be presented to aviation and ground system combat developers on how the system can be used to mitigate these forces and reduce further harm to the casualty during transport. The research study will be used to provide guidance to combat developers and requirements branches as UAV/UGV casualty extraction doctrine is being developed. Develop a commercialization plan identifies the potential of the product to save casualty lives during rapid extraction from a hostile environment to a safe location to be further treated. The commercialization plan needs to include development pathways and military potential, to also include civilian sectors.

REFERENCES:

1. U.S. Army Publication: Medical Evacuation ATP 4-02.2 http://armypubs.army.mil/doctrine/DR_pubs/dr_a/pdf/atp4_02x2.pdf

2. Article by Andrew Tarantola: Why the Human Body Can’t Handle Heavy Acceleration. http://gizmodo.com/why-the-human-body-cant-handle-heavy-acceleration-1640491171

3. USAF Flight Surgeons Guide: https://www.google.com/url?sa=t&rct=j&q=&esrc=s&source=web&cd=5&ved=0CDgQFjAEahUKEwiFxpHrn-rHAhVH0oAKHYirAmY&url=http%3A%2F%2Fwww.dlielc.edu%2Folc%2Fset%2FANC%2Favdocs%2Findex.php%3Fdir%3D3%2BAerospace%2BMedicine%252F%26download%3DUSAF%2BFlight%2BSurgeons%2BGuide.pdf&usg

4. OPNAVINST 3710.7U NATOPS General Flight and Operating Instructions. http://www.med.navy.mil/sites/nmotc/nami/arwg/Documents/WaiverGuide/OPNAVINST_3710_7U_General_NATOPS.pdf

KEYWORDS: combat casualty care, UAV, Casualty evacuation, human flight limitations, G-Forces and altitude limitations

TECHNOLOGY AREA(S): Biomedical

OBJECTIVE: Develop and characterize novel, flexible live-attenuated tetravalent vaccines against dengue capable of rapid adaptation to natural antigenic variation and generation of a dengue serotype-specific humoral cellular responses.

DESCRIPTION: Dengue viral (DENV) diseases is the third most significant natural infectious threat to U.S. deployed forces (Burnette et al., 2008). An efficient vaccine against DENV is urgently needed to protect our servicemen and women when they deploy to DENV endemic regions across the globe such as South East Asia, Southern Asia, the Middle East, Latin America, South America, and Africa. Dengue fever and dengue hemorrhagic fever are two disease syndromes associated with infection by one of the four serotypes of DENV, and these are highly debilitating diseases – impacting mission efficiency for 14-28 days. (Gibbons et al, 2012). Transmission to troops occurs through the bite of the Aedes aegypti or the Aedes albopictus mosquito (Heinz et al. 2012). These mosquitoes are traditionally found in many theaters of current and future US military operations. DENV has affected US troops since the Spanish American War and there are in field examples from modern conflicts of how DENV can have a marked effect on troop health. For example, in operations Restoring Hope (Somalia, 1993) and Uphold Democracy (Haiti, 1994), 20-30% and of hospitalized febrile troops were positive for dengue fever (Trofa et al., 1997).

Live vaccines are typically the most efficacious, as compared to inactivated and subunit vaccines. Since live attenuated vaccines (LAV) activate all branches of the immune system, they have been proven to be superior in terms of immune response and protection when compared to inactivated or subunit vaccines. However, two major problems have impeded the development of a live DENV vaccine:

1. Low stability of attenuated phenotype in previous vaccine candidates. Traditional LAV development has been an empirical procedure, relying on a lengthy serial passage adaptation in non-human tissue culture cells and/or experimental animals. The resulting virus may acquire mutations that mute its virulence when inoculated into the human host; however, the number of mutations responsible for the attenuated phenotype is usually very small (less than 10), often resulting in unacceptably low genetic stability of the phenotype (Shimizu et al., 2004).

2. Unbalanced immune response. The difficulty of DENV vaccine development is compounded by the presence of four distinct serotypes and the phenomenon of Antibody-Dependent Enhancement (ADE) (Murphy, et al., 2011). Each DENV serotype is sufficiently different, such that there is no long term cross-protection between serotypes, and epidemics caused by multiple serotypes (hyper-endemicity) can result. It is believed that when four fully competent dengue viruses are introduced at the same time that they potentially interfere with each other and/or replicate with different efficiency. This potentially leads to an unbalanced immune response perhaps leading short lived cross protective antibodies and longer term sub neutralizing antibodies. ADE occurs when prior infection with one serotype predisposes an individual to an enhanced severity of disease (dengue hemorrhagic fever/dengue shock syndrome, DHF/DSS) upon re-infection with a different serotype (Thomas and Endy, 2011). During ADE, antibodies against the first virus bind, but do not neutralize the second virus, instead increasing its infectivity by Fc receptor-mediated uptake into a wider variety of host cells. Thus, it is essential that a successful DENV vaccine induces an adequate and equal level of protection against all four serotypes simultaneously, or else the incomplete vaccine response against one of the serotypes, may predispose an incompletely immunized vaccine recipient to ADE. Lastly, natural antigenic variation among the four serotypes of Dengue necessitates a Dengue vaccine that could be rapidly and rationally modified to respond to these shifts as opposed to one that would require the standard empirical process used for traditional live vaccines.

Ideally a dengue vaccine for our deploying troops should be: 1) easily administered with immunity in 28 days; 2) offer durable/long-term immunity; 3) protect/seroconvert against ALL serotypes, 4) significantly reduce risk of ADE; and 5) allow for rapid scale-up of production.

PHASE I: Proposed efforts should be focused on the development of a tetravalent, live attenuated dengue vaccine– ideally a vaccine that induces both a humoral and cellular dengue-specific response. The approach used for dengue vaccine development should be easily applied to rapidly generate other live attenuated vaccines of interest to the Army as the need arises. Activities in Phase I should include generation and characterization of lead candidate(s) for the tetravalent dengue vaccine. At the conclusion of Phase I work, successful offerors must present in vitro and if possible within the time constraints in vivo characterization of a lead candidate(s) for at least 2 components of a tetravalent, live attenuated dengue vaccine including attenuation, immunogenicity, and manufacturing stability.

PHASE II: Phase II work should focus on 1) identifying and producing candidate for all 4 dengue virus serotypes 2) develop a tetravalent dengue product(s) 3) evaluate candidates in vivo and in vitro as demonstrated in phase 1 to include demonstrating balanced replication among the 4 serotypes 4) manufacture non-GMP lots 5) demonstrate efficacy of the lead candidate(s) in non-human primates, including quantification of antibody response in vaccinated NHPs as well as vaccine strain induction of an anti-dengue T cell response Selection of dose and dosing schedule should be addressed in NHP studies. Durability and level of immunity must be demonstrated in challenge studies for both monovalent and tetravalent vaccine formulations. Protection must be demonstrated in tetravalent vaccine using established NHP challenge experiments.

PHASE III DUAL USE APPLICATIONS: Produce vaccine under GMP pilot production conditions, conduct an FDA phase 1 trial in healthy adults of monovalent and/or tetravalent formulations. Safety and immunogenicity must be explored during the clinical trial including durability with a minimum 6 month post vaccination time point. The results of the clinical trial will permit an evaluation of immune response and preliminary safety assessment. Vaccine shelf-life assessment will also be initiated during this phase. Provide a comprehensive report of all testing performed during this phase, including a complete searchable dataset of the clinical trial. In addition potential exploitation of this platform to other viral vaccine needs can be explored during this phase. These might include Chikungunya or Zika.

REFERENCES:

1. Burnette WN, Hoke CH Jr, Scovill J, Clark K, Abrams J, Kitchen LW, Hanson K, Palys TJ, Vaughn DW. Infectious diseases investment decision evaluation algorithm: a quantitative algorithm for prioritization of naturally occurring infectious disease threats to the U.S. military. Mil Med. 2008 Feb;173(2):174-81

2. Gibbons, Robert V., et al. "Dengue and US military operations from the Spanish-American War through today." Emerg Infect Dis 18.4 (2012): 623-630

3. Heinz, F.X., Stiasny, K., 2012. Flaviviruses and flavivirus vaccines. Vaccine 30, 4301-4306

4. Murphy, B.R., Whitehead, S.S., 2011. Immune response to dengue virus and prospects for a vaccine. Annu Rev Immunol 29, 587-619

5. Shimizu, H., Thorley, B., Paladin, F.J., Brussen, K.A., Stambos, V., Yuen, L., Utama, A., Tano, Y., Arita, M., Yoshida, H., Yoneyama, T., Benegas, A., Roesel, S., Pallansch, M., Kew, O., Miyamura, T., 2004. Circulation of type 1 vaccine-derived poliovirus in the Philippines in 2001. J Virol 78, 13512-13521

6. Thomas, S.J., Endy, T.P., 2011. Critical issues in dengue vaccine development. Curr Opin Infect Dis 24, 442-450

7. Trofa AF, DeFraites RF, Smoak BL, Kanesa-thasan N, King AD, Burrous JM,MacArthy PO, Rossi C, Hoke CH Jr. Dengue fever in US military personnel in Haiti.JAMA. 1997 May 21;277(19):1546-8

KEYWORDS: Infectious diseases, Dengue, Dengue vaccines, Live attenuated dengue vaccines, Tetravalent dengue vaccines, Antibody dependent enhancement

TECHNOLOGY AREA(S): Biomedical

OBJECTIVE: Develop novel methods to engineer vascularized tissues for volumetrically large organs.

DESCRIPTION: In the last decade, research groups have made great advances in stem cell science, tissue engineering, biomedical engineering, and other areas of regenerative medicine that have begun to show results. However, the wealth of basic research advances has thus far produced few clinically applicable solutions to organ injury and disease, especially in complex organs such as the heart, liver, lungs, and kidneys. The ability to vascularize thick tissue constructs has been identified as a key limiting factor in this effort (3,4,5).

Currently, technology to create thick tissue assays and constructs that closely recapitulate human physiology is limited by the capacity for diffusion of nutrients into the engineered tissue. This limits our capabilities to creating tissues that are no more than a few millimeters thick. Recent advances have begun to develop technologies capable of expanding this to greater than 1 cm thick tissues (3,7,8); however, more work is required to create sustainable constructs capable of use in drug testing, disease modeling, and for tissue repair and regeneration in human patients. Of particular interest in this SBIR topic is the capability of producing thick tissue replacements for patients in need of repair or regeneration of lost tissue and functionality of the major thick-walled organs including: heart, liver, lung, kidneys, pancreas, or muscle tissues. Functional evaluations should be appropriate to the organ type selected and are not limited to, but may include:

Organ: Cell Types: Functional Evaluation:

The HHS-led Vision 2020 report estimates that the world market for replacement organ therapies alone is over $350 billion. In addition, the Alliance for Regenerative Medicine’s annual report states that “potential savings from regenerative medicine treatments – for the U.S. … have been estimated at approximately $250 billion a year.” However, clinical translation of actual solutions has been limited and slow to progress beyond a few, relatively non-complex organs such as the trachea or bladder (2, 3). This solicitation aims to extend that capability to more complex, thick-tissue organs that could be useful for injured warfighters, in disaster response, and for the health-care industry at large.

PHASE I: The performer will develop concepts for a technology capable of engineering and maintaining a thick tissue construct (>1cm) with parenchymal cells functioning similar to those of one of the thick-walled organs including the heart, lung, liver, kidneys, or muscles and demonstrate feasibility of the technology concepts through in-vitro or ex-vivo testing. Feasibility will be established through demonstration of tissue construction, perfusion, and limits on cell necrosis of the tissue over a minimum one week using an in-vitro or ex-vivo system.

PHASE II: The performer will test the long-term survival and functionality of in-vitro tissue constructs and demonstrate a minimum 30 day trial period noting levels of tissue necrosis and demonstrate structural and functional performance parameters relative to those of a healthy adult’s performance of the same tissue type. Performance metrics of a healthy adult must be included in the final comparison and must include the structural and functional performance measurements appropriate for the organ model system selected. The model selected should be appropriate to the tissue type being tested. Engineered vascular tissues must be evaluated in an in vivo setting where the tissue is able to recapitulate the intended function of the native tissue. Demonstrations where the engineered vascular tissue is able to fully replace the function of the native tissue over a period of several days are encouraged but not required.

PHASE III DUAL USE APPLICATIONS: Vascular engineering to support manufacturing of complex biological tissues is an open problem with a large potential market and with direct applicability across the full spectrum of medical treatment, biological manufacturing, diagnostics, on-chip drug screening, and long-term unattended biologically based sensor platforms. The effort should include a description of plans to address the commercialization of the underlying technology. Potential paths to commercialization may benefit from potential future funding under programs administered through USAMRMC such as USAMMDA or CDMRP. It must describe one or more specific Phase III military applications and/or supported S&T or acquisition programs as well as the most likely path for transition of the SBIR from research to operational capability. For example, the proposal might relate the use of cryopreservation solutions, protocols or equipment to the potential use in the treatment of particular diseases or conditions of military interest. Specific defense applications include manufacture of engineered tissues. Additionally, the Phase III section must include (a) one or more potential commercial applications OR (b) one or more commercial technologies that could be potentially inserted into defense systems as a result of this particular SBIR project. The performer or a suitable partner will pursue development of the approach to permit the vascular engineering of successively larger tissues and organs. This award mechanism will bridge the gap between laboratory-scale innovation and entry into a recognized FDA regulatory pathway leading to commercialization.

REFERENCES:

1. ASSAY and Drug Development Technologies, Ryan, S-L., A-M. Baird, G. Vaz, A.J. Urquhart, M. Senge, D.J. Richard, K.J. O’Byrne, and M. Davies Anthony. February 2016, 14(1): 19-28. doi:10.1089/adt.2015.670

2. The Promise and Potential of ‘Organs-on-Chips’ as Preclinical Models, Session Moderator: A. Bahinski. Participants: R. Horland, D. Huh, C. Mummery, D.A. Tagle, and T. MacGill. Applied In Vitro Toxicology. December 2015, 1(4): 235-242. doi:10.1089/aivt.2015.29002.rtl

3D bioprinting of tissues and organs, S. V. Murphy, A. Atala, Nature Biotechnology, vol 32, no. 8, August 2014

4. Vascularization strategies for tissue engineering, Tissue Engineering Part B, vol 15, no 3, Sept 2009 (http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2817665/)

5. Vascularization is the key challenge in tissue engineering, Advanced Drug Delivery Review, April 30, 2011 (http://www.ncbi.nlm.nih.gov/pubmed/21396416)

6. Microvascular Guidance: a challenge to support the development of vascularized tissue engineering construct, I Sukmana, Scientific World Journal, 2012. (http://www.ncbi.nlm.nih.gov/pubmed/22623881)

3D Bioprinting of Vascularized, Heterogeneous Cell-Laden Tissue Constructs, D.B. Kolesky, R.L. Truby, A.S. Gladman, T.A. Busbee, K.A. Homan, J. A. Lewis, Advanced Materials, Volume 26, Issue 19, pages 3124–3130, May 21, 2014

8. Three Dimensional Bioprinting of Thick Vascularized Tissues, D. B. Kolesky, K. A. Homan, M. A. Skylar-Scott, J. A. Lewis, Proceedings of the National Academy of Science, vol. 113 no. 12, March 22, 2016

KEYWORDS: vascular, tissue, engineered, trauma, regenerative, medicine

A17-069

TITLE: Novel Concentration Technology