ABSTRACT
BACKGROUND: The current biomarkers alpha-fetoprotein and human chorionic gonadotropin have limited sensitivity and specificity for diagnosing malignant germ-cell tumours (GCTs). MicroRNAs (miRNAs) from the miR-371-373 and miR-302/367 clusters are overexpressed in all malignant GCTs, and some of these miRNAs show elevated serum levels at diagnosis. Here, we developed a robust technical pipeline to quantify these miRNAs in the serum and cerebrospinal fluid (CSF). The pipeline was used in samples from a cohort of exclusively paediatric patients with gonadal and extragonadal malignant GCTs, compared with appropriate tumour and non-tumour control groups. METHODS: We developed a method for miRNA quantification that enabled sample adequacy assessment and reliable data normalisation. We performed qRT-PCR profiling for miR-371-373 and miR-302/367 cluster miRNAs in a total of 45 serum and CSF samples, obtained from 25 paediatric patients. RESULTS: The exogenous non-human spike-in cel-miR-39-3p and the endogenous housekeeper miR-30b-5p were optimal for obtaining robust serum and CSF qRT-PCR quantification. A four-serum miRNA panel (miR-371a-3p, miR-372-3p, miR-373-3p and miR-367-3p): (i) showed high sensitivity/specificity for diagnosing paediatric extracranial malignant GCT; (ii) allowed early detection of relapse of a testicular mixed malignant GCT; and (iii) distinguished intracranial malignant GCT from intracranial non-GCT tumours at diagnosis, using CSF and serum samples. CONCLUSIONS: The pipeline we have developed is robust, scalable and transferable. It potentially promises to improve clinical management of paediatric (and adult) malignant GCTs.
Subject(s)
Biomarkers, Tumor/blood , Central Nervous System Neoplasms/diagnosis , MicroRNAs/blood , Neoplasm Recurrence, Local/diagnosis , Neoplasms, Germ Cell and Embryonal/diagnosis , Ovarian Neoplasms/diagnosis , Testicular Neoplasms/diagnosis , Adolescent , Biomarkers, Tumor/cerebrospinal fluid , Carcinoma, Embryonal/blood , Carcinoma, Embryonal/cerebrospinal fluid , Carcinoma, Embryonal/diagnosis , Central Nervous System Neoplasms/blood , Central Nervous System Neoplasms/cerebrospinal fluid , Child , Child, Preschool , Choriocarcinoma, Non-gestational/blood , Choriocarcinoma, Non-gestational/cerebrospinal fluid , Choriocarcinoma, Non-gestational/diagnosis , Chorionic Gonadotropin/blood , Chorionic Gonadotropin/cerebrospinal fluid , Endodermal Sinus Tumor/blood , Endodermal Sinus Tumor/cerebrospinal fluid , Endodermal Sinus Tumor/diagnosis , Female , Germinoma/blood , Germinoma/cerebrospinal fluid , Germinoma/diagnosis , Humans , Infant , Infant, Newborn , Male , MicroRNAs/cerebrospinal fluid , Neoplasm Recurrence, Local/blood , Neoplasm Recurrence, Local/cerebrospinal fluid , Neoplasms, Germ Cell and Embryonal/blood , Neoplasms, Germ Cell and Embryonal/cerebrospinal fluid , Ovarian Neoplasms/blood , Ovarian Neoplasms/cerebrospinal fluid , Polymerase Chain Reaction , Sacrococcygeal Region , Sensitivity and Specificity , Testicular Neoplasms/blood , Testicular Neoplasms/cerebrospinal fluid , alpha-Fetoproteins/cerebrospinal fluid , alpha-Fetoproteins/metabolismSubject(s)
Brain Neoplasms/pathology , Carcinoma in Situ/pathology , Germinoma/pathology , Neoplasms, Multiple Primary/pathology , Testicular Neoplasms/pathology , Brain Neoplasms/blood , Brain Neoplasms/cerebrospinal fluid , Carcinoma in Situ/blood , Carcinoma in Situ/cerebrospinal fluid , Chorionic Gonadotropin, beta Subunit, Human/blood , Chorionic Gonadotropin, beta Subunit, Human/cerebrospinal fluid , Fatal Outcome , Germinoma/blood , Germinoma/cerebrospinal fluid , Humans , Male , Neoplasm Proteins/blood , Neoplasm Proteins/cerebrospinal fluid , Neoplasms, Multiple Primary/blood , Neoplasms, Multiple Primary/cerebrospinal fluid , Testicular Neoplasms/blood , Testicular Neoplasms/cerebrospinal fluid , Young Adult , alpha-Fetoproteins/analysis , alpha-Fetoproteins/cerebrospinal fluidABSTRACT
Ascending myelopathy developed in a previously irradiated 10-year-old boy after intraventricular methotrexate and cytosine arabinoside were given for central nervous system relapse of acute lymphoblastic leukemia. The course was fatal in 10 weeks. Cerebrospinal fluid myelin basic protein levels, indicating a demyelinative process, rose prior to the onset of clinical symptoms and remained at very high levels until death. Myelin basic protein may be useful as a predictor of chemotherapy-associated neurotoxicity.
Subject(s)
Leukemia, Lymphoid/pathology , Myelin Basic Protein/cerebrospinal fluid , Spinal Cord Neoplasms/secondary , Antineoplastic Combined Chemotherapy Protocols/therapeutic use , Asparaginase/administration & dosage , Child , Combined Modality Therapy , Cytarabine/administration & dosage , Humans , Hydrocortisone/administration & dosage , Injections, Spinal , Leukemia, Lymphoid/cerebrospinal fluid , Leukemia, Lymphoid/therapy , Male , Methotrexate/administration & dosage , Prednisone/administration & dosage , Radiotherapy Dosage , Recurrence , Spinal Cord Neoplasms/cerebrospinal fluid , Spinal Cord Neoplasms/pathology , Testicular Neoplasms/cerebrospinal fluid , Testicular Neoplasms/pathology , Testicular Neoplasms/secondary , Vincristine/administration & dosageABSTRACT
Serum values of alpha-fetoprotein (AFP) and human chorionic gonadotrophin (HCG) have been used to monitor disseminated testicular carcinoma. Serial measurements of these markers have been used to monitor the response to therapy, to follow the progress of disease, and to detect subclinical recurrences. With increasingly effective chemotherapy for systemic disease, central nervous system (CNS) metastases in testicular carcinoma are becoming increasingly important as a cause of treatment failure. Cerebrospinal fluid (CSF) values of AFP and HCG seem to be important ancillary acids in the neurosurgical management of CNS metastases from testicular cancer. Our preliminary experience with three cases suggests that these CSF markers (plus computerized tomograhic scanning) should be evaluated in patients with this disease.
