Making tissue-specific progenitor cells that possess the ability to survive, but have not passed through the pluripotency state is a highly desirable goal of regenerative medicine. The technique known as “direct reprogramming” uses various genetic tricks to transdifferentiate mature, adult cells into different cell types that can be used for regenerative treatments.
Juan Carlos Izpisua Belmonte and his colleagues from the Salk Institute for Biological Studies in La Jolla, California and his collaborators from Spain have used direct reprogramming to convert human skin cells into a type of white blood cells.
These experiments began with harvesting skin fibroblasts from human volunteers that were then forced to overexpress a gene called “Sox2.” The Sox2 gene is heavily expressed in mice whose bone marrow stem cells are being reconstituted with an infusion of new stem cells. Thus this gene might play a central role is the differentiation of bone marrow stem cells.
Sox2 overexpression in human skin fibroblasts cause the cells express a cell surface protein called CD34. Now this might seem so boring and unimportant, but it is actually really important because CD34 is expressed of the surfaces of hematopoietic stem cells. Hematopoietic stem cells make all the different types of white and red blood cells in our bodies. Therefore, the expression of these protein is not small potatoes.
In addition to the expression of CD34, other genes found in hematopoietic stem cells were also induced, but not strongly. Thus overexpression of SOX2 seems to induce an incipient hematopoietic stem cell‐like status on these fibroblasts. However, could these cells be pushed further?
Gene profiling of hematopoietic stem cells from Umbilical Cord Blood identified a small regulatory RNA known as miR-125b as a factor that pushes SOX2-generated CD34+ cells towards an immature hematopoietic stem cell-like progenitor cell that can be grafted into a laboratory animal.
When SOX2 and miR-125b were overexpressed in combination, the cells transdifferentiated into monocytic lineage progenitor cells.
What are monocytes? They are a type of white blood cells and are, in fact, the largest of all white blood cells. Monocytes compose 2% to 10% of all white blood cells in the human body. They play multiple roles in immune function, including phagocytosis (gobbling up bacteria and other stuff), antigen presentation (identifying and altering other cells to the presence of foreign substances), and cytokine production (small proteins that regulate the immune response).
Monocytes express a molecule on their cell surfaces called CD14, and when human fibroblasts overexpressed Sox2 and miR-125b, they became CD14-expressing cells that looked and acted like monocytes. These cells were able to gobble up bacteria and other foreign material, and when transplanted into a laboratory animal, these directly reprogrammed cells generated cells that established the monocytic/macrophage lineage.
Cancer patients, and other patients with bone marrow diseases can have trouble making sufficient white blood cells. A technique like this can generate transplantable monocytes (at least in laboratory animals) without many of the drawbacks associated with reprogramming human cells into hematopoietic stem cells that possess true clinical potential. Also because this technique skips the pluipotency stage, it is potentially safer.