Experienced Postdoctoral Researcher with a demonstrated history of working with genetically modified immune cells. Skilled in Genome Editing/Crispr, Multi-color Flow Cytometry, Molecular Biology, Cell Culture and Immunotherapy.
Bachelor of Science, Sabanci University (2009)
Master of Science, University Of London (2011)
Doctor of Philosophy, Sabanci University (2017)
Post-doctoral research associate, Nova Southeastern University, Immunology/Cell Therapy (2019)
PhD, Sabanci University, Molecular Immunology (2017)
Master of Science, King's College London, Immunology (2010)
Bachelor of Science, Sabanci University, Biological Sciences and Bioengineering (2009)
Maria Grazia Roncarolo, Postdoctoral Faculty Sponsor
Engineering antigen-specific NK cell lines against the melanoma-associated antigen tyrosinase via TCR gene transfer
EUROPEAN JOURNAL OF IMMUNOLOGY
2019; 49 (8): 1278–90
Introduction of Chimeric Antigen Receptors to NK cells has so far been the main practical method for targeting NK cells to specific surface antigens. In contrast, T cell receptor (TCR) gene delivery can supply large populations of cytotoxic T-lymphocytes (CTL) targeted against intracellular antigens. However, a major barrier in the development of safe CTL-TCR therapies exists, wherein the mispairing of endogenous and genetically transferred TCR subunits leads to formation of TCRs with off-target specificity. To overcome this and enable specific intracellular antigen targeting, we have tested the use of NK cells for TCR gene transfer to human cells. Our results show that ectopic expression of TCR α/β chains, along with CD3 subunits, enables the functional expression of an antigen-specific TCR complex on NK cell lines NK-92 and YTS, demonstrated by using a TCR against the HLA-A2-restricted tyrosinase-derived melanoma epitope, Tyr368-377 . Most importantly, the introduction of a TCR complex to NK cell lines enables MHC-restricted, antigen-specific killing of tumor cells both in vitro and in vivo. Targeting of NK cells via TCR gene delivery stands out as a novel tool in the field of adoptive immunotherapy which can also overcome the major hurdle of "mispairing" in TCR gene therapy.
View details for DOI 10.1002/eji.201948140
View details for Web of Science ID 000478643000013
View details for PubMedID 31054264
Successive crystal structure snapshots suggest the basis for MHC class I peptide loading and editing by tapasin
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
2019; 116 (11): 5055–60
MHC-I epitope presentation to CD8+ T cells is directly dependent on peptide loading and selection during antigen processing. However, the exact molecular bases underlying peptide selection and binding by MHC-I remain largely unknown. Within the peptide-loading complex, the peptide editor tapasin is key to the selection of MHC-I-bound peptides. Here, we have determined an ensemble of crystal structures of MHC-I in complex with the peptide exchange-associated dipeptide GL, as well as the tapasin-associated scoop loop, alone or in combination with candidate epitopes. These results combined with mutation analyses allow us to propose a molecular model underlying MHC-I peptide selection by tapasin. The N termini of bound peptides most probably bind first in the N-terminal and middle region of the MHC-I peptide binding cleft, upon which the peptide C termini are tested for their capacity to dislodge the tapasin scoop loop from the F pocket of the MHC-I cleft. Our results also indicate important differences in peptide selection between different MHC-I alleles.
View details for DOI 10.1073/pnas.1807656116
View details for Web of Science ID 000460911500051
View details for PubMedID 30808808
View details for PubMedCentralID PMC6421438
Functional Assessment for Clinical Use of Serum-Free Adapted NK-92 Cells
2019; 11 (1)
Natural killer (NK) cells stand out as promising candidates for cellular immunotherapy due to their capacity to kill malignant cells. However, the therapeutic use of NK cells is often dependent on cell expansion and activation with considerable amounts of serum and exogenous cytokines. We aimed to develop an expansion protocol for NK-92 cells in an effort to generate a cost-efficient, xeno-free, clinical grade manufactured master cell line for therapeutic applications. By making functional assays with NK-92 cells cultured under serum-free conditions (NK-92SF) and comparing to serum-supplemented NK-92 cells (NK-92S) we did not observe significant alterations in the viability, proliferation, receptor expression levels, or in perforin and granzyme levels. Interestingly, even though NK-92SF cells displayed decreased degranulation and cytotoxicity against tumor cells in vitro, the degranulation capacity was recovered after overnight incubation with 20% serum in the medium. Moreover, lentiviral vector-based genetic modification efficiency of NK-92SF cells was comparable with NK-92S cells. The application of similar strategies can be useful in reducing the costs of manufacturing cells for clinical use and can help us understand and implement strategies towards chemically defined expansion and genetic modification protocols.
View details for DOI 10.3390/cancers11010069
View details for Web of Science ID 000457233300033
View details for PubMedID 30634595
View details for PubMedCentralID PMC6356567