| dc.description.abstract | The aim of this thesis was to establish the therapeutic value of the IFITM1
monoclonal antibodies and to design and develop therapeutically valuable
recombinant monoclonal antibodies so as to study the implication of these
novel antibodies in cancer therapy. Cancer metastasis is one of the main
interests that has given rise to the design and development of innovative
strategies for cancer therapeutics. The Interferon Induced Transmembrane
Protein 1(IFITM1), a notable member of the IFITM family of proteins has
been identified as one of the most up-regulated trans-membrane proteins in
metastatic breast cancer and cervical adenocarcinoma. This interferon-regulated
protein is also involved in cell migration, invasion in glioma and
squamous cancers. This PhD aimed to study IFITM1 as a pro-invasive cancer
target by the use of IFITM1 monoclonal antibodies that were raised against the
extracellular domain of the human IFITM1 gene. The epitope mapping of
IFITM1 revealed the binding activity of the IFITM1 monoclonal antibody.
This gave the opportunity to design and generate to new IFITM1-specific
molecular tools, in the form of recombinant IFITM1 targeted murine scFv
antibody, IFITM1-CPG2 yeast fusion protein antibody for potential
application in ADEPT as well as a Mouse-Human Chimeric IFITM1 antibody
secreting mammalian cell line. The immunohistochemical staining of IFITM1
in tissue micro array from breast, colon and oeosphegal cancer has revealed
that the majority of these cancers produce this protein. However, IFITM1 is
over produced in cervical cancer indicating it’s selective over expression in
cervical cells. This PhD endeavored to investigate the expression of IFITM1 at
a translational and transcriptional level and to study the clinical significance of
IFITM1 in cervical cancer. The antibody dependent cell mediated cytotoxic
activity of the chimeric IFITM1 antibody was found to be cytotoxic to SiHa cells in vitro. In the future these molecular tools could be used to regulate and
further characterize the activity of this transmembrane protein antibody. In an
effort to better understand the mechanisms that regulate the activity and the
over production of the IFITM1 gene and its interacting proteins, a proteomic
screen of cervical cancer cells was carried out using data-independent
SWATH-MS on an AB SCIEX TripleTOF™ mass spectrometer. This Mass Spec
analysis provided us with a host of IFITM1 biomarkers and revealed that the
IFITM1 gene and its binding proteins also cross link with the IRF1 pathway.
The data presented in this thesis, demonstrates that the IFITM1 gene can be
targeted to either stimulate or inhibit IFITIM1 signaling to engage IFITM1 as
a potential pro-invasive extracellular receptor as a target in antibody cancer
therapy.
In summary, this thesis aimed to confirm the activity and the binding
specificity of the IFITM1 antibody. Additionally, this thesis demonstrated a
promising application of the recombinant antibody in the ADEPT technology.
Characterization of IFITM1 mAb effector functions indicated that the
antibody was cytotoxic to cervical cancer cells. This highlights an important
element in the immune suppressive tumour microenvironment. And finally,
this thesis also provides the basis for the production of recombinant mouse
human chimeric antibodies that are a part of a new group of
immunotherapeutic molecules paving the way for cancer therapeutics. | en |
