Epigenetically Enhanced PDT (ePDT) Induces Significantly Higher Levels of Multiple Extrinsic Pathway Apoptotic Factors than Standard PDT Resulting in Greater Extrinsic and Overall Apoptosis of CTCL

摘要

Aminolevulinate-based photodynamic therapy (ALA-PDT) selectively eliminates diseased tissues primarily through the induction of intrinsic apoptotic pathway. ALA-PDT is a first-line therapy for actinic keratosis, however, it is less effective for cutaneous T-cell lymphoma (CTCL). We have previously demonstrated that the resistance of CTCL to apoptosis correlates with decreased expression of death receptors such as FAS, and that methotrexate functions as an epigenetic regulator that re-establishes the susceptibility of CTCL to extrinsic pathway apoptosis. We showed previously that MTX augments the effectiveness of PDT by sensitizing cells to apoptosis by induction of apoptotic factors, a process we call “epigenetically enhanced” PDT (ePDT). Here, in CTCL cell lines, leukemic CTCL cells and normal blood T cells, we analyzed multiple components of the FAS, TRAIL and TNF families using multispectral imaging of immunostained cytopreparations, a quantitative technique with 5-fold greater sensitivity than standard immunocytology. ePDT induced significantly greater FAS, FASL, TRAIL-R1 & -R2, and TNFα levels than standard PDT. This correlated with significantly greater induction of extrinsic pathway apoptosis and/or overall apoptosis in all CTCL samples. There was no appreciable effect on normal T cells. These data set the stage for clinical trials of ePDT as a novel localized treatment of CTCL.