Biomedical subjects

Deniz Dalkara

Publications and source records attributed to Deniz Dalkara.

4 recordsLinked to original sources

PubMed2026

Interindividual variability in immune response to AAV ocular gene delivery across species impedes immunomonitoring.

Adeno-associated viruses (AAVs) have been used in gene therapy, especially for inherited retinal diseases. Despite their effectiveness in gene transduction, immune responses to the AAV capsid and transgene products have been reported, which can compromise both the efficacy and the safety of AAV-mediated therapies. The eye is regarded as an immune-privileged organ where immune activity is constitutively suppressed. Here, we highlight that immunomonitoring in an ocular gene transfer reveals variable immune responses, whatever the species (human clinical trial, nonhuman primates, mice), the site of injection, the cassette, and the dose. We further explored factors contributing to this variability, investigating the potential correlation among immune parameters in a controlled experimental setting. In a syngeneic murine model after a subretinal injection of AAV, our results highlight an interindividual variability of immune parameters, emphasizing the importance of considering inherent variability among individuals when designing personalized therapies.

Animals↗

PubMed2006

Intracellular protein delivery with a dimerizable amphiphile for improved complex stability and prolonged protein release in the cytoplasm of adherent cell lines.

Direct delivery of functionally active proteins into cells represents an emerging strategy for laboratory investigation and therapeutic applications. For this purpose, we developed a novel amphiphile (CholCSper) consisting of cholesterol linked to carboxy-spermine by a cysteine. This amphiphile is dimerizable upon mild oxidation of the thiol to disulfide and it was used in formulation with DOPE to prepare an intracellular protein delivery system. The stabilization of the CholCSper assemblies by chemical conversion of CholCSper into its gemini amphiphile afforded the production of homogeneous assemblies with proteins whose sizes are easier to control. Furthermore, the cholesterol moiety has an effect on the density of the complexes formed with proteins and leads to a prolonged protein release in the cytoplasm of cells exposed to the protein carrier assemblies.

Animals↗

PubMed2006

Cationic oligonucleotide-peptide conjugates with aggregating properties enter efficiently into cells while maintaining hybridization properties and enzymatic recognition.

Oligonucleotide delivery is a crucial issue for therapeutical purposes and is often addressed by conjugation to short cationic peptides although with controversial results. To further examine this mechanism, a 15-mer anionic oligonucleotide was conjugated to a cationic peptide in order to obtain a diblock compound with an overall positive charge with aggregation properties. These microaggregates were efficiently internalized in cells via the expeditious pathway used by commercial gene delivery systems. Moreover, stability of the duplex formed with the complementary sequence increased without inhibiting oligonucleotide enzyme recognition as shown by the properties of the conjugate to prime chain elongation by Taq DNA polymerase in a linear amplification/sequencing process.

Amino Acid Sequence↗

PubMed2004

Intracytoplasmic delivery of anionic proteins.

Protein delivery is emerging as an interesting alternative to gene delivery. We have used our experience with transfection to develop a technique for efficient delivery of anionic proteins such as antibodies into the cytoplasm of cells. As for DNA, when complexed with cationic lipids, large amounts of proteins are shown to enter adherent cells via ubiquitously expressed syndecans. However, protein surface area rather than electric charge ratio governs the delivery characteristics. Delivery of anti-beta-actin and anti-alpha-tubulin IgG's leads to fiber depolymerization. Intracellular delivery of an antibody could thus be regarded as another method for interfering with gene activity.

Animals↗