This Small Business Innovation Research (SBIR) Phase I project will develop advanced methods to temporarily reprogram hematopoietic stem cells (HSCs) that will enable ex vivo production of blood in quantities suitable for transfusion. A transfection-free method will be used that can be easily translated into clinical use. Every two seconds someone in the U.S. needs blood and more than 38,000 blood donations are needed every day. In order for ex vivo blood production to be clinically relevant, the expansion cell culture must be able to produce red blood cells in the range contained in a unit of blood: 2.5x1012cells. There exists a fundamental obstacle in reaching this number of cells. CD34+(HSCs) and erythroid progenitors are typically only capable of short-term, or restricted ex vivo expansion. The main objective of this project is to reprogram CD34+(HSCs) such that cellular mechanisms are activated to force cellular proliferation. Despite research efforts from around the world, reaching clinical significant quantities of blood in culture has not been previously possible. The goal is to produce clinically relevant quantities of blood and create a shift in clinical practice paradigms for the treatment of the anemic patient as well as the blood banking industry. What are the broader impacts of the proposed activity?
The broader impact/commercial potential of this project, if successful, will be to create a viable alternative to allogenic blood transfusion, and compete with traditional blood banking and the use of recombinant erythropoietin (EPO). We expect that successful development of this technique will have a high societal impact on safety and availability of blood for transfusions as well as a reduction in hemolytic transfusion reactions. Although the safety of transfusions has improved markedly there is still a non-zero probability of getting HIV (1:1.8 Million), or Hepatitis C (1:1 Million). This work is expected to significantly enhance the scientific and technological understanding of ex vivo production of blood and provide a pathway to translation into clinical medicine. This project will have a significant impact in the $20 billion global blood transfusion market, as well as the EPO market, which is valued at about $53 Billion.
. Reporting Period: 07/01/2013 - 12/31/2013 This Small Business Innovation Research Phase I project demonstrated the successful production of engineered blood in the laboratory from human adult stem cells present in a small venous blood sample. We utilized specific specialized proteins that can directly and temporarily control gene expression, along with cell growth factors. We were able to produce 100,000,000 red blood cells that were grown in the laboratory from the small number of adult stem cells that are found within a small sample of human adult blood. Adult stem cells are a type of stem cell that is present within each tissue type in your body and remain present in scant quantities for the duration of your life. Stem cell research and tissue engineering efforts have predominantly focused on the use of embryonic stem cells which are derived from a fetus. Although these embryonic stem cells are relatively easy to grow in the laboratory and can be differentiated into many different tissue types, there are significant ethical concerns associated with using and obtaining cells from a fetus. The field of Regenerative Medicine is reaching a road block over the use of embryonic stem cells and the ability to translate tissue engineering work into FDA approved uses for patients. Virus induced gene transfer into cells that can induce stem cell like properties is a common technique in the field of Regenerative Medicine, but gene transfer can induce cancer and represents another road block the field of Regenerative Medicine. This SBIR project overcomes both of these obstacles. This approach was found to be successful as a low-cost manufacturing method, and easily translatable into clinical medicine. One of the key limiting factors in the in-vitro production of blood today is the high cost of reagents and the need to start with large amounts of stem cells generated through either umbilical cord blood or embryonic stem cells. This approach has been shown to be viable using peripheral blood with simple extraction techniques. In this project, we increase the adult stem cell proliferation efficiency by altering cellular mechanisms without gene manipulation. We impact the field by demonstrating the usefulness of adult stems cells in blood production. This will lessen the need for donated blood and improve the safety of the blood supply. This project also demonstrated the feasibility of in-vitro blood production that could be used in place of traditional donated blood. There are widespread societal benefits to this work. Due to demographic changes, it is expected that the US will not have enough blood to satisfy demand by 2020 using only donated blood. There are additional benefits such as the ability to produce large quantities of type O negative, which can supply people universally and reduce the potential for mistakes in blood typing as well as improvements in safety due to reduced potential for pathogens, and transfusion reactions.
