There is an art to science, and science in art.
Isaac Asimov

Current projects

Over the past two decades, stem cell biology and cell culture methods have greatly improved. The industrial production of transfusion products has become a practical reality. A major focus of the lab is generating red blood cells from induced pluripotent stem cells for use as diagnostic tools, drug delivery vehicles and tranfusions for patients with allo-imunization.

Large scale production of cultured red blood cells

The goal of this project is to develop a cost-effective method for producing large numbers of RBCs starting from iPSCs.

We have developed the chemically-defined Robust Erythroid Differentiation (RED) protocol to differentiate iPSCs into RBCs.

The RED protocol is highly efficient since it yields more than 100,000 cells per iPSC.

the RED protocol is economical because it does not require any albumin and only small amounts of transferrin because supplementation with an iron chelator, allows transferrin recycling to take place in cell culture.

Because most applications of red blood cell culture require very large number of cells, continuing to decrease production costs is essential to bring the technology to the clinic.

To decrease costs further we are developing iPSC lines containing constitutive mutations in cytokines receptors and signal transduction pathways. Upon differentiation, expression of these mutations allows the production of RBCs from iPSCs without the use of cytokines during most of the culture period.

A novel therapy for Thrombotic Thormbocytopenic Purpura

The use of red blood cells as drug delivery vehicles is one of most promising application of red blood cell culture technology

The main objective of this project is to evaluate the feasibility of using red blood cells (cRBCs) that express GPI-anchored ADAMTS13, as a treatment for Thrombotic Thrombocytopenic Purpura (TTP).

The project has two aims: (1) to produce cRBCs that are resistant to auto-antibodies responsible for idiopathic TTP, (2) to test the effectiveness of GPI-ADAMTS13-cRBCs in compensating ADAMTS13 loss in a mouse model.

If successful, these experiments will provide proof-of-concept for using engineered cRBCs as a novel treatment for TTP and validate a powerful platform for producing and testing therapeutic iPSC-derived cRBCs.

Gene therapy for congenital erythroid disorders

Patients with congenital erythroid disorders have a variety of mutations, making it difficult to develop a gene therapy intervention that can treat all cases.

There are three main approaches for gene therapy for hemoglobinopathies under investigation. Gene addition is generalizable but has disadvantages such as variable expression levels, the presence of toxic endogenous genes, and a risk of insertional mutagenesis. Gene editing is precise but challenging to apply to disorders caused by multiple mutations and is not applicable to large deletions. BCL11a enhancer editing is very promising but specific for a subtype of β-hemoglobinopathies.

We are developing a method to introduce therapeutic transgenes in human primary CD34+ cells in a site specific manner. This method provides exciting opportunities for gene therapy, but raises the question of what the ideal "safe harbor" would be.

We are testing the idea that the β-globin cluster is a good safe harbor due to its location in a gene-poor region and its well-understood transcriptional regulation.

We are optimizing our knock-in strategy, characterizing the properties of the β-cluster safe harbor, and attempting to demonstrate the correction of globin expression in CD34+ cells from patients with sickle cell disease and α- and β-thalassemia.

This novel approach to mutation-agnostic knock-in should increase the number of patients eligible for gene therapy and decrease the cost of developing treatments for patients with atypical genotypes.