Investigating the Microbiome and Volatile Organic Compound Profile of Children With Neuroblastoma

Investigating the Microbiome and Volatile Organic Compound Profile of Children With Neuroblastoma - a Pilot Study

Background: Malignant tumors may lead to a catabolic state with loss of muscle and adipose tissue. The full picture of catabolism is termed cachexia and is associated with significant morbidity and mortality of cancer patients. Although the full picture is rarely observed up to 50% of children with cancer suffer from significant malnourishment. Additionally to tumor-induced catabolism, side-effects of chemotherapy may be problematic for the patients. In this regard up to 60% of children suffer from gastrointestinal mucositis presenting with nausea, vomiting, diarrhea or constipation and abdominal pain. In the worst case, mucositis may lead to bacterial translocation with life-threatening inflammatory response. Clinically this may require a reduction of the dosage or the number of chemotherapy cycles resulting in reduced effectivity. Up to now the therapy of mucositis is only symptomatic. Recent research of the applicant has shown a significant reduction of Lactobacilli in mice with neuroblastoma (a malignant childhood tumor). The dysbiosis was associated with catabolism, increased gut permeability and inflammation. Astonishingly, chemotherapy alone also leads to a significant reduction of Lactobacilli compared to sham mice, which may be linked to the development of mucositis clinically. Overall, the intestinal microbiome seems to play an essential role in the development of tumor-associated catabolism and chemotherapy-induced mucositis.

Aim: The aim of this project is to determine if the changes in the intestinal microbiome observed in mice can also be seen in children with neuroblastoma.

Methods: One part of the study will include 10 children with neuroblastoma (inclusion after verification of the diagnosis) and 10 healthy controls. The fecal microbiome will be determined by 16S-ribosomal deoxyribonucleic acid (rDNA) pyrosequencing. Volatile organic compounds in the breath will be sampled and measured by Gas Chromatography/Mass Spectroscopy. A basic science human work package will address the question if there are differences.

In the second part serial investigations in children with neuroblastoma will assess whether or not these patients show alterations of the intestinal microbiome under chemotherapy.

Study Overview

• Status: Recruiting
• Conditions: Microbial Colonization; Neuroblastoma; Children, Only
• Intervention / Treatment: Diagnostic test: Initial fecal microbiome; Diagnostic test: Initial fecal volatile organic compounds; Diagnostic test: Initial breath volatile organic compounds; Diagnostic test: Microbiome under chemotherapy; Diagnostic test: Fecal volatile organic compounds under chemotherapy; Diagnostic test: Breath volatile organic compounds under chemotherapy; Diagnostic test: Final microbiome; Diagnostic test: Final fecal volatile organic compounds; Diagnostic test: Final breath volatile organic compounds

• Study Type: Interventional
• Enrollment (Anticipated): 20
• Phase: Not Applicable

Contacts and Locations

• Study Contact: Name: Christoph Castellani, MD; Phone Number: 80217 +43/316/385; Email: christoph.castellani@medunigraz.at
• Study Contact Backup: Name: Georg Singer, MD; Phone Number: 83722 +43/316/385; Email: georg.singer@medunigraz.at

Study Locations

• Austria
  • Styria
    • Graz, Styria, Austria, 8036
      • Recruiting
      • Department of Paediatric and Adolescent Surgery, Medical University of Graz, Austria
      • Contact: Christoph Castellani, MD; Phone Number: 80217 +43/316/385; Email: christoph.castellani@medunigraz.at
      • Sub-Investigator: Daniela Sperl, MD
      • Sub-Investigator: Georg Singer, MD

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: 1 year to 6 years (Child)
• Accepts Healthy Volunteers: No
• Genders Eligible for Study: All

Description

Inclusion Criteria:

• Age 2-8 years
• Neuroblastoma group: verified neuroblastoma
• Control group: absence of pulmonary or gastro-intestinal disease
• Written parental informed consent obtained

Exclusion Criteria:

• Active gastro-intestinal or pulmonary disease
• Antibiotic or probiotic treatment within 3 weeks before sampling
• Negative parental informed consent

Study Plan

How is the study designed?

Design Details

• Primary Purpose: Basic Science
• Allocation: Non-Randomized
• Interventional Model: Parallel Assignment
• Masking: None (Open Label)

Number of Arms

2

Arms and Interventions

Participant Group / Arm	Intervention / Treatment
Experimental: Neuroblastoma group; 10 children with neuroblastoma. Inclusion after verification of diagnosis and informed consent. Sampling of fecal microbiome (Initial microbiome, microbiome under chemotherapy, final microbiome), fecal volatile organic compounds (initial fecal volatile organic compounds, fecal volatile organic compounds under chemotherapy and final fecal volatile organic compounds) and breath organic volatile compounds (initial breath organic compounds, breath volatile organic compounds under chemotherapy and final breath volatile organic compounds). Samples will be taken after verifying diagnosis before initiation of chemotherapy, 1 week after completion of each cycle and 3 weeks after the end of chemotherapy.	Diagnostic test: Initial fecal microbiome; Stool sampling for fecal microbiome analysis by 16S rDNA pyrosequencing. Neuroblastoma group and Control group.; Diagnostic test: Initial fecal volatile organic compounds; Volatile organic compound analysis of stool samples by gas chromatography/mass spectroscopy Neuroblastoma group and Control group.; Diagnostic test: Initial breath volatile organic compounds; Breath sampling for organic compound analysis by gas chromatography/mass spectroscopy Neuroblastoma group and Control group.; Diagnostic test: Microbiome under chemotherapy; Stool sampling under chemotherapy of children in neuroblastoma group (1 sample 1 week after completion of each chemotherapy cycle). Chemotherapy according to Société Internationale d´Onclogie Pediatrique Neuroblastoma Group (SIOPEN) guidelines; Diagnostic test: Fecal volatile organic compounds under chemotherapy; Stool sampling under chemotherapy of children in neuroblastoma group (1 sample 1 week after completion of each chemotherapy cycle). Chemotherapy according to SIOPEN guidelines. Neuroblastoma group; Diagnostic test: Breath volatile organic compounds under chemotherapy; Breath sampling under chemotherapy of children in neuroblastoma group (1 sample 1 week after completion of each chemotherapy cycle). Chemotherapy according to SIOPEN guidelines. Neuroblastoma group; Diagnostic test: Final microbiome; Stool sampling 3 weeks after completion of chemotherapy Neuroblastoma group; Diagnostic test: Final fecal volatile organic compounds; Stool sampling 3 weeks after completion of chemotherapy Neuroblastoma group; Diagnostic test: Final breath volatile organic compounds; Breath sampling 3 weeks after completion of chemotherapy Neuroblastoma group
Other: Control group; 10 children without gastro-intestinal or pulmonary disease as age and sex matched controls to the neuroblastoma group. Patients will be recruited from paediatric surgery. Inclusion after informed consent. Sampling of fecal microbiome (initial fecal microbiome), fecal volatile organic compounds (initial fecal volatile organic compounds) and breath organic volatile compounds (initial breath volatile organic compounds). Samples will be taken as age and sex matched controls for the neuroblastoma group. Sampling will be done once after obtaining informed consent.	Diagnostic test: Initial fecal microbiome; Stool sampling for fecal microbiome analysis by 16S rDNA pyrosequencing. Neuroblastoma group and Control group.; Diagnostic test: Initial fecal volatile organic compounds; Volatile organic compound analysis of stool samples by gas chromatography/mass spectroscopy Neuroblastoma group and Control group.; Diagnostic test: Initial breath volatile organic compounds; Breath sampling for organic compound analysis by gas chromatography/mass spectroscopy Neuroblastoma group and Control group.

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Difference of alpha and beta diversity, relative abundance of fecal bacteria at different levels (phylum, class, order, family and genus levels) between neuroblastoma and control group	Alpha and beta diversity, relative bacterial abundance at different levels in percent.	Neuroblastoma group: within 48h after diagnosis, before initiation of chemotherapy. Control group: within 24h after obtaining informed consent.
Change of alpha and beta diversity, relative abundance of fecal bacteria at different levels (phylum, class, order, family and genus levels) under chemotherapy in the neuroblastoma group	Alpha and beta diversity, relative bacterial abundance at different levels in percent.	Within 48h after diagnosis, before initiation of chemotherapy; 1 week after each chemotherapy cycle and 3 weeks after the end of chemotherapy.

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Difference of anthropometric data between neuroblastoma and control group.	Body weight (in kg) and height (in m) will be determined to calculate the body mass index (BMI in kg/m^2).	Neuroblastoma group: within 48h after diagnosis. Control group: within 24h after obtaining informed consent.
Change of anthropometric data under chemotherapy in the neuroblastoma group	Body weight (in kg) and height (in m) will be determined to calculate the Body mass index (BMI in kg/m^2).	Within 48h after diagnosis, before initiation of chemotherapy; 7 days after completion of each chemotherapy cycle and 3 weeks after the end of chemotherapy.
Change of mucositis score under chemotherapy in the neuroblastoma group.	Assessment of the mucositis score according to the WHO criteria (WHO handbook for reporting results of cancer Treatment; WHO Offset publication no 48) The score contains 5 subitems which are evaluated separately. At the end a total score is derived by adding the results of all items. Subitem 1: oral mucosa; range 0 (best) to 4 (worst) Subitem 2: nausea and vomiting; range from 0 (best) to 4 (worst) Subitem 3: diarrhea; range from 0 (best) to 4 (worst) Subitem 4: constipation; range from 0 (best) to 4 (worst) Subitem 5: abdominal pain; range from 0 (best) to 4 (worst)	Within 48h after diagnosis, before initiation of chemotherapy; 7 days after completion of each chemotherapy cycle and 3 weeks after the end of chemotherapy.
Difference of breath volatile organic compounds between neuroblastoma and control group.	Volatile organic compounds in ppb in the exhaled breath.	Neuroblastoma group: within 48h after diagnosis. Control group: within 24h after obtaining informed consent.
Difference of stool volatile organic compounds between neuroblastoma and control group.	Volatile organic compounds in ppb in stool samples.	Neuroblastoma group: within 48h after diagnosis. Control group: within 24h after obtaining informed consent.
Change of breath volatile organic compounds under chemotherapy in the neuroblastoma group.	Volatile organic compounds in ppb in the exhaled breath.	Within 48h after diagnosis, before initiation of chemotherapy; 1 week after each chemotherapy cycle and 3 weeks after the end of chemotherapy.
Change of stool volatile organic compounds under chemotherapy in the neuroblastoma group.	Volatile organic compounds in ppb in stool samples.	Within 48h after diagnosis, before initiation of chemotherapy; 1 week after each chemotherapy cycle and 3 weeks after the end of chemotherapy.

Collaborators and Investigators

• Sponsor: Medical University of Graz
• Investigators: Principal Investigator: Christoph Castellani, MD, Department of Paediatric and Adolescent Surgery, Medical University of Graz, Austria

Study record dates

Study Major Dates

• Study Start (Actual): May 7, 2018
• Primary Completion (Anticipated): June 30, 2022
• Study Completion (Anticipated): December 31, 2022

Study Registration Dates

• First Submitted: May 7, 2018
• First Submitted That Met QC Criteria: May 22, 2018
• First Posted (Actual): June 4, 2018

Study Record Updates

• Last Update Posted (Actual): February 25, 2020
• Last Update Submitted That Met QC Criteria: February 24, 2020
• Last Verified: February 1, 2020

More Information

Terms related to this study

• Keywords: Microbiome; Neuroblastoma; Volatile organic compounds
• Additional Relevant MeSH Terms: Pathologic Processes; Neoplasms by Histologic Type; Neoplasms; Neoplasms, Glandular and Epithelial; Disease Attributes; Neoplasms, Neuroepithelial; Neuroectodermal Tumors; Neoplasms, Germ Cell and Embryonal; Neoplasms, Nerve Tissue; Neuroectodermal Tumors, Primitive; Neuroectodermal Tumors, Primitive, Peripheral; Infections; Communicable Diseases; Neuroblastoma
• Other Study ID Numbers: 0001 (Cancer Research Institute)

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: No

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No