Blood sample collection from Met-pa

Peripheral whole blood samples of 7.5 mL were collected from 52 Met-pa and 3 healthy volunteers after informed consent (Guthrie Clinic IRB#1808–45). All of the experiments were done in accordance with the U.S Federal Policy for the Protection of Human Subjects and approved by the Institutional Review Board of the Guthrie Clinic. The participants in this study were fully informed regarding the objective of the current study and written consent was obtained. Of these Met-pa, 48 and 44 samples were used to investigate the correlation between the initial CTC and CAF levels with cancer prognosis. Furthermore, 30 patient samples were collected through respective chemotherapy regimens at the following time points: (i) Baseline (pre-chemotherapy treatment), (ii) Following 1 cycle of chemotherapy and (iii) Following 2 cycles of chemotherapy. Finally, 12 patients were used as negative controls to fully determine the efficacy of our TRAIL-liposomal therapy. De-identified blood samples were shipped from the Guthrie Clinic to Vanderbilt University and processed within 24 h. Figure 1 and Table 1 show information related to the patient group that participated in this study. During blood collection, patients received the same chemotherapy regimen established at the baseline time point. In addition, the disease progression was determined by computerized tomography (CT scan) at each time point, where the patients were grouped as stable disease, disease progression and deceased. Stable disease is defined as when the metastatic burden has not changed. However, disease progression refers to patients displaying a growth of an existing metastatic lesion or the appearance of a new lesion in a distant organ. Patients that did not survive the cancer treatment were grouped as deceased (1–9 months of survival). After 1 year of blood collection, the clinical status of the patients analyzed in this study was determined and recorded. The patients were classified into three different categories: stable disease, progression and deceased patients. As stated above, stable disease and progression was determined using CT scan. However, when patients died, the date was recorded to determine the overall survival of this cohort of patients. To determine the effect of chemotherapy on CTC and CAF counts, patients that showed stable disease throughout chemotherapy treatment were considered for this part of the study. In this part, we wanted to investigate the CTC/CAF mobilization by treatment rather than by natural cancer progression.

Diagram that display the blood collection from Met-pa and their clinical background information. a Blood samples were collected from a total of 52 patients diagnosed with a spectrum of cancer types at metastatic stage, including: renal, prostate, pancreatic, gastric, esophageal, colorectal, ovarian, endometrial, cervical, breast and lung carcinoma. From these 52 patients, 26 were followed through chemotherapy treatment. This figure was created by the authors for this article

Clinical background information of the Met-pa participating in this study