Insulin-like growth factor-1 and neurotrophin-3 gene therapy prevents motor decline in an X-linked adrenoleukodystrophy mouse model

X-linked adrenoleukodystrophy (X-ALD) is the most common inherited peroxisomal disorder characterized by a progressive demyelination of the central nervous system. The marked loss of myelin and oligodendrocytes observed in the disease prompted us to evaluate the therapeutic potential of insulin-like growth factor-1 and neurotrophin-3, two potent inducers of myelin formation and oligodendrocyte survival. Viral vectors engineered to produce insulin-like growth factor-1 or neurotrophin-3 were administrated into the cerebrospinal fluid of an X-linked adrenoleukodystrophy mouse model. We show that viral-based, long-lasting delivery of insulin-like growth factor-1 and neurotrophin-3 significantly halts the progression of the disease and leads to potent protective effect against the demyelination process

Presence of Gal-alpha1,3Gal epitope on xenogeneic lines: implications for cellular gene therapy based on the encapsulation technology

Patrick Aebischer

Exposure to human serum induces the lysis of xenogeneic cells through natural antibodies and complement activation. The carbohydrate Galactose-alpha1,3-Galactose (Gal-alpha1,3-Gal) epitope, has been shown to be the principal antigenic determinant on target cells. This reaction is, therefore, particularly important for xenogeneic cell-based therapy. As a first step toward the evaluation of the impact of this phenomenon for encapsulated xenogeneic cells, we have evaluated the presence of the Gal-alpha1,3Gal epitope on two cell lines currently being used for the systemic delivery of protein in the periphery or the treatment of neurodegenerative diseases. In the second part of the study, we have tested and compared human serum and cerebrospinal fluid (CSF) for the presence of xenoreactive natural antibodies (XNAs) and their potential impact on the survival of xenogeneic cells. Fluorescence-activated cell sorting analysis indicated that baby hamster kidney (BHK) cells expressed low levels of the alpha-Gal epitope, whereas mouse myoblast C2C12 cells were extensively stained with the specific IB4-lectin. There was a direct correlation between serum killing and the level of Gal-alpha1,3-Gal epitope expression on these cells. Importantly, we showed that CSF did not lyse BHK and C2C12 cells as determined by cytotoxic crossmatch assays. The reaction was specific as the addition of soluble Gal-alpha1,3-Gal sugar to human serum effectively reduced cell killing, and the overproduction of alpha-1,3-galactosyltransferase in BHK cells significantly increased inactivation by human serum. To interfere with this antibody-antigen reaction and develop cell lines particularly suitable for cell-based therapy, we either selected C2C12 clones expressing low levels of Gal-alpha1,3-Gal or high levels of alpha-1,2-fucosyltransferase. These cells were found to be resistant to complement-mediated cytolysis. These strategies may, therefore, protect encapsulated xenogeneic cells transplanted in the periphery or the central nervous system even in an unlikely event of a blood-brain barrier breakage and the post-transplantation development of an antibody response.

2003

Parkinson's Disease: Gene Therapies

Patrick Aebischer, Philippe Guillaume Coune, Bernard Schneider

With the recent development of effective gene delivery systems, gene therapy for the central nervous system is finding novel applications. Here, we review existing viral vectors and discuss gene therapy strategies that have been proposed for Parkinson's disease. To date, most of the clinical trials were based on viral vectors to deliver therapeutic transgenesto neurons within the basal ganglia. Initial trials used genes to relieve the major motor symptoms caused by nigrostriatal degeneration. Although these new genetic approaches still need to prove more effective than existing symptomatic treatments, there is a need for disease-modifying strategies. The investigation of the genetic factors implicated in Parkinson's disease is providing precious insights in disease pathology that, combined with innovative gene delivery systems, will hopefully offer novel opportunities for gene therapy interventions to slow down, or even halt disease progression.

Cold Spring Harbor Lab Press, Publications Dept2012

Intracerebroventricular Injection of Adeno-Associated Virus 6 and 9 Vectors for Cell Type-Specific Transgene Expression in the Spinal Cord

Patrick Aebischer, Elisabeth Anne Dirren, Bernard Schneider, Veronica Setola, Christopher Towne

In the context of motoneuron diseases, gene delivery as an experimental or therapeutic approach is hindered by the challenge to specifically target cell populations that are widely distributed along the spinal cord. Further complicating the task, transgenes often need to be delivered to motoneurons and/or glial cells to address the non-cell-autonomous mechanisms involved in disease pathogenesis. Intracerebroventricular (ICV) injection of recombinant adeno-associated viruses (AAVs) in newborn mice allows distributing viral vectors throughout the central nervous system while limiting undesired transduction of peripheral organs. Here, we show that by combining the appropriate set of AAV serotype and promoter, specific transgene expression can be achieved in either motoneurons or astrocytes along the whole mouse spinal cord. ICV injection of recombinant AAV6 with the cytomegalovirus (cmv) promoter preferentially targets motoneurons, whereas AAV9 particles combined with the astrocyte-specific gfaABC(1)D promoter lead to significant transgene expression selectively targeted to astrocytes. Importantly, ICV coinjection of both AAV6-cmv and AAV9-gfaABC(1)D results in segregated expression of two different transgenes in motoneurons and astrocytes, respectively. Relevance of viral vector delivery via the cerebrospinal fluid was further investigated in young nonhuman primates. Intracisternal injection of recombinant AAV6-cmv led to robust cervical transduction of motoneurons, highlighting the potential of this approach for gene therapy and modeling of motoneuron diseases.

Mary Ann Liebert2014

Presence of Gal-alpha1,3Gal epitope on xenogeneic lines: implications for cellular gene therapy based on the encapsulation technology

Patrick Aebischer

2003