Photodynamic therapy: Theoretical and experimental approaches to dosimetry.

摘要

Singlet oxygen (1O2) is the major cytotoxic species generated during photodynamic therapy (PDT), and 1O 2 reactions with biological targets define the photodynamic dose at the most fundamental level. We have developed a theoretical model for rigorously describing the spatial and temporal dynamics of oxygen (3O 2) consumption and transport and microscopic 1O 2 dose deposition during PDT in vivo. Using experimentally established physiological and photophysical parameters, the mathematical model allows computation of the dynamic variation of hemoglobin-3O 2 saturation within vessels, irreversible photosensitizer degradation due to photobleaching, therapy-induced blood flow decrease and the microscopic distributions of 3O2 and 1O 2 dose deposition under various irradiation conditions. mTHPC, a promising photosensitizer for PDT, is approved in Europe for the palliative treatment of head and neck cancer. Using the theoretical model and informed by intratumor sensitizer concentrations and distributions, we calculated photodynamic dose depositions for mTHPC-PDT. Our results demonstrate that the 1O 2 dose to the tumor volume does not track even qualitatively with long-term tumor responses. Thus, in this evaluation of mTHPC-PDT, any PDT dose metric that is proportional to singlet oxygen creation and/or deposition would fail to predict the tumor response. In situations like this one, other reporters of biological response to therapy would be necessary. In addition to the case study of mTHPC-PDT, we also use the mathematical model to simulate clinical photobleaching data, informed by a possible blood flow reduction during treatment. In a recently completed clinical trial at Roswell Park Cancer Institute, patients with superficial basal cell carcinoma received topical application of 5-aminolevulinic acid (ALA) and were irradiated with 633 nm light at 10-150 mW cm-2 . Protoporphyrin IX (PpIX) photobleaching in the lesion and the adjacent perilesion normal margin was monitored by fluorescence spectroscopy. We successfully simulate the in vivo photobleaching of PpIX in this patient population over a wide range of irradiances using the PDT model. For most cases, the rate of bleaching slows as treatment progresses, leaving a fraction of the PpIX unbleached despite sustained irradiation. To account for this feature, the model predicts that incorporation of ALA-PDT-induced blood flow reduction is necessary. In addition to using the theoretical method to understand the dose deposited by photodynamic therapy, experimentally, we propose a potential dose metric for Pc 4-PDT. Pc 4 is a promising second generation photosensitizer that is now in Phase I clinical trials for the treatment of cutaneous lesions. We have observed a significant irradiation-induced increase in Pc 4 fluorescence in tumor cell monolayers. The amount of the fluorescence increase observed in vitro strongly correlates to the cell death and mitochondrial swelling reported by the clonogenic cell survival assay and light scattering measurements, respectively. Based on those biological responses, we anticipate that irradiation-induced fluorescence enhancement in Pc 4-PDT may be a potential dose metric.