Identification and Characterization of Predictive Factors of Onset of Bone Metastases in Cancer Patients

MATERIALS AND METHODS: 1) Removal of whole blood from peripheral vein and separation of plasma from enrolled metastatic cancer patients; 2) Isolation of tumor exosomes circulating from the plasma of cancer patients (Kit Invitrogen: Total Exosome Isolation Reagent); 3) Next generation sequencing methods (NGS) will be used to evaluate the expression profiles of miRNAs isolated from circulating tumor exosomes (MiSeq Illumina System); 4) Mass spectrometry methods will be used for the proteome analysis of circulating exosomes isolated from the plasma of cancer patients (Triple TOFF 5600 Plus System AND HPLC analysis of bioactive and biomarker peptides); 5) 3D cultures for the study of cross-talk between circulating tumor exosomes (isolated from the patients' plasma) and bone tissue cells (using commercial cell lines) as part of the metastatic process (Cellec perfusion bioreactor BioteK AG).

Enrollment Procedures: Patients considered eligible will be enrolled in the study, after providing a written informed consent.

Data Collection: Clinical data will be retrieved by patient's source document. A protocol-specific CRF reporting the results of the analyses will be provided. A CRF is required and should be completed for each included subject.

Ethics: The clinical trial protocol and its documents will be sent before initiating the study to the competent Authorities and Ethics Committees of each participating country for its approval. The responsible investigator will ensure that this study is conducted in agreement with either the most updated Declaration of Helsinki and all the international and local laws that apply to clinical trials and to patient protection. The protocol has been written, and the study will be conducted according to the principles of the ICH Harmonized Tripartite Guideline for Good Clinical Practice (ref: http://www.emea.eu.int/pdfs/human/ich/013595en.pdf).

Informed Consent: All patients will be informed, by the investigator, of the aims of the study, the possible risks and benefits that will derive from the study participation. The Investigator must clearly inform that the patient is free to refuse participation in the study and that can withdraw consent at any time and for any reason. They will be informed as to the strict confidentiality of their patient data, but that their medical records may be reviewed for trial purposes by authorized individuals other than their treating physician. The informed consent procedure must conform to the ICH guidelines on Good Clinical Practice. This implies that "the written informed consent form should be signed and personally dated by the patient or by the patient's legally acceptable representative".