Safety of CD34+ Hematopoietic Stem Cells and CD4+ T Lymphocytes Transduced with LVsh5/C46 in HIV-1 Infected Patients with High-Risk Lymphoma.

Although the risk of developing lymphoma has decreased in the highly active antiretroviral therapy era, this cancer remains the major cause of mortality in HIV-infected patients. Autologous hematopoietic stem cell transplantation (ASCT) outcome does not differ for HIV-infected versus HIV-uninfected patients. We propose to develop a new treatment for HIV-associated high-risk lymphoma based on autologous transplantation of two genetically modified products: CD4+ T lymphocytes and CD34+ hematopoietic stem cells (HSPCs). The cells will be transduced ex vivo with the Cal-1 lentiviral vector encoding for both a short hairpin RNA (shRNA) against CCR5 (sh5) and the HIV-1 fusion inhibitor C46. The transduced cells will be resistant to HIV infection by two complementary mechanisms: impaired binding of the virus to the cellular CCR5 co-receptor and decreased fusion of the virus as C46 interacts with gp41 and inhibits HIV infection. This phase I/II pilot study, also entitled GENHIV, will involve two French participating centers: Saint Louis Hospital and Necker Hospital in Paris. We plan to enroll five HIV-1-infected patients presenting with high-risk lymphoma and require a treatment with ASCT. The primary objective of this study is to evaluate the safety, feasibility, and success of engraftment of Cal-1 gene-transduced CD4+ T lymphocytes and CD34+ HSPCs.

Industrial approach in developing an advanced therapy product for bone repair.

Mesenchymal stem cells (MSCs) are multipotent cells with therapeutic applications. The aim of our work was to develop an advanced therapy product for bone repair, associating autologous human adipose-derived MSCs (ASCs) with human bone allograft (TBF; Phoenix). We drew up specifications that studied: (a) the influence of tissue collection procedures (elective liposuction or non-invasive resection) and patient age on cell number and function; (b) monolayer cell culture conditions and osteodifferentiation and particularly the possibility of reducing stages of culture; and (c) the bone construct preparation and especially the comparison between two types of cells seeded on bone allograft (number of cultured processed lipoaspirate (PLA) cells and monolayer-expanded ASCs) and cultured for 1, 2 and 3 weeks. The results showed that tissue harvesting techniques and patient age did not affect PLA cell number and ASC cloning efficiency. PLA cells can be directly osteodifferentiated (instead of culturing them in expansion medium first and then differentiating them) and these cells were able to mineralize when they were cultured in an osteogenic medium containing calcium chloride. PLA cells directly seeded on bone allograft for a minimum of 3 weeks of culture in this osteogenic medium expressed osteocalcin and colonized the matrix better than monolayer-expanded ASCs. This work detailed the specifications of a pharmaceutical laboratory to develop an advanced therapy product and this current approach is promising for bone repair.