SOW.doc

NIH Clinical Center

Critical Care Medicine Department CCMD research section Statement of Work

BACKGROUND INFORMATION

The Critical Care Medicine Department (CCMD) was established in November 1977 to provide medical care for critically ill patients throughout the Clinical Center who have potentially reversible medical problems. Critical care medicine may be defined as the observation, diagnosis, treatment, and rehabilitation of patients with overt or potential failure of vital functions.

The Department is currently composed of the following sections:

· Clinical Medical Services

· Critical Care Therapy and Respiratory Care

· Research Section

· AIDS Section

· Administrative Services

CCMD Mission To provide high quality and cost effective clinical service that meets the needs of CC patients and NIH Institutes, to facilitate the research goals of NIH Institutes and investigators, and to conduct high quality research and training in Critical Care Medicine.

CCMD Research Section

The Critical Care Medicine Department conducts active research programs in bedside clinical investigation, research in animal models, and molecular biology.

Full-time research activities commence at the beginning of the second year of the fellowship for fellows interested in the critical care subspecialty alone. For those interested in dual board eligibility, their second-year program is individually designed and consists of clinical rotations required for board eligibility in the particular subspecialties. After completion of second subspecialty training, full-time research activities commence. Major topics of research interest within the Department include:

· The immunopathogenesis of septic shock

· Studies of immune cellular function in respiratory distress syndrome, asthma, and other forms of severe pulmonary disease

· Biomarkers and novel therapies in pulmonary arterial hypertension

· Biomarkers of acute cellular rejection in heart transplantation

· Diagnosis and treatment of opportunistic infections

· Studies of immune cellular function and therapy of AIDS

· Pathogenesis and treatment of anthrax

· Emerging infectious diseases

· Biology and clinical effects of nitric oxide

· Acute lung injury

· Pulmonary arterial hypertension

· Role as transcription regulatory factor

· Functional genomics of critical illness

· Human acute inflammation

· Animal model of septic shock and multiple organ failure

· Pulmonary arterial hypertension

· Solid organ transplantation

· Emerging Infectious Disease

PURPOSE

The purpose of this SOW is to request a PO to obtain analysis of the metabolomic responses in serum from human volunteers who have been challenged with intravenous endotoxin. This study was done under a collaborative research agreement between Duke University, the Foundation for NIH and the NIH Clinical Center. The samples are unique as they reflect a Phase I study that evaluated the dose response of a new preparation of Escherichia coli (E. coli group O 113:H10:K negative) endotoxin in healthy human volunteers. Four different doses of reference endotoxin were tested in four cohorts of 6 patients each subjects (2 subjects saline placebo and 4 subjects given either 0.6, 1, 2, and 4 ng/kg body weight endotoxin). These data will provide unique information that reflects the initial responses of humans to a non-infectious bacterial product that mimics the initial stages of a bacterial infection. This has importance as primary research information as well as reference data for other investigators using this product in their research. Seven time points (baseline,1, 2, 3, 4, 6, 8 hrs) will be analyzed for each subject (161 total samples available)

REQUIREMENTS

Vendor will provide global metabolomic profiling of 161 human plasma samples by the following methods:

· Vendor shall be able to survey a wide range of very polar to non-polar compounds (typically 400-1000) extracted from as little as 50 μl of biofluid (human plasma).

· Extracted samples are to be split into five aliquots for different chromatography and MS platforms, three UHPLC-MS methods and one hydrophilic interaction chromatography (HILIC)-MS, with one aliquot held in reserve; LC-MS samples are spiked with 11 standards at known concentrations.

· These four chromatography and MS systems complement each other in the range of biochemicals measured, providing broad coverage of metabolic space within each sample.

· For compounds observed on multiple platforms, the platform with the best analytical characteristics (e.g. fewest interfering peaks or highest signal to noise) is to be generally used for the analysis of that compound.

· Vendor shall have a spectral library of over 4100 standard authentic metabolites and over 7000 ―unnamed previously observed metabolites.

· Vendor must provide a suite of software packages to automatically integrate each ion across retention time and then uses that ionic information, which may include additional MS/MS fragmentation information and retention time, to identify the compound. After a compound is identified in a sample, one of the characteristic and higher abundance ions is used to determine a relative concentration of that compound in each sample. Day to day instrument variation can be addressed through simple scaling of multiday means, but requires the samples to be randomized across the run days for each experiment.

· Other QC steps include addition of an internal standard (human plasma), a pure water blank, and a solvent blank for each assay plate. Finally, when the software has finished analyzing the sample, all of the data is loaded into a visual user interface that allows curation of the data for QC purposes, leading to inclusion of only those compounds with the highest degree of confidence in the final data set.

SCOPE OF WORK

We have performed a dose escalation study of IV endotoxin in 24 volunteers studied in 4 cohorts of 6 patients (2 controls and 4 endotoxin challenged subjects in each group; endotoxin doses given 0.6, 1, 2, 4 ng/kg). Serial blood samples were obtained over the first 8 hours and at 24 hours.

Seven time points (baseline,1, 2, 3, 4, 6, 8 hrs) will be analyzed for each subject (161 total samples available)

QUALIFICATIONS

Vendor must meet the following requirements:

· Extract and measure 400-1000 named metabolites from as little as 50 μl of biofluid (human plasma).

· Run extraction across four (4) different chromatography and MS platforms, three UHPLC-MS methods and one hydrophilic interaction chromatography (HILIC)-MS,

· For compounds observed on multiple platforms, the platform with the best analytical characteristics is to be generally used for the analysis of that compound.

· Shall have a spectral library of over 4100 standard authentic metabolites and over 7000 ―unnamed previously observed metabolites.

· Provide a suite of software packages to automatically integrate each ion across retention time and then uses that ionic information, which may include additional MS/MS fragmentation information and retention time, to identify the compound.

· After a compound is identified in a sample, one of the characteristic and higher abundance ions is used to determine a relative concentration of that compound in each sample.

· QC steps include addition of an internal standard (human plasma), a pure water blank, and a solvent blank for each assay plate.

· Data is loaded into a visual user interface that allows curation of the data for QC purposes, leading to inclusion of only those compounds with the highest degree of confidence in the final data set.

OBJECTIVES

Our current analysis of these samples include: 1) sequencing of mRNA and miRNA in whole blood at 3 time points, 2) analysis of secreted miRNA in blood, 3) proteomic analysis using Somalogic aptamers. A metabolomic profile with the dose escalation would be a useful complement to our other analysis.

PERIOD OF PERFORMANCE

June 2017-May 2018; Analyze the 161 samples and will send final data and results to NIH within 10-14 weeks of the date that all 161 samples are received in acceptable condition.

Management and Resources Plan

REPORTING REQUIREMENTS

The contractor shall make information systems available for NIH to develop monthly status reports if needed.

DELIVERABLES

Analysis and Deliverable: The samples proceed through a multi-step process including analysis using four different platforms: UHPLC-MS/MS(+ESI) UPHLC-MS/MS(-ESI), HILIC-MS (early positive ionization), and UPHLC-MS/MS (late positive ionization).

Identification: Each detected biochemical is automatically compared to a reference standard in our database, using retention index and mass spectrum. The result is a broad range of biochemicals identified including amino acids, carbohydrates, lipids, nucleic acids and cofactors. Report back the mass of the quant ion along with what platform the metabolite was detected on (e.g., LC-MS NEG, LC-MS POS) and its retention index.

Quantitation: Once biochemicals have been identified, the project manager analyzes the data to check for quality and then applies statistical analysis to determine which biochemicals show significant changes. p and q values to determine and identify significant biochemical changes Results: mView deliverable provides information on a broad range of biochemical, and how they change in response to experimental variables. The information is delivered to the client in a variety of ways, at a number of different levels to provide a complete picture of the biochemical effects.

Interpretation: One page summary highlighting biochemical differences. Short presentation slide deck providing graphical support for highlighted biochemical differences.

Place of performance

All the work will be performed within the state of North Carolina and in the city of Morrisville.

NON-PERSONAL SERVICE STATEMENT:

Contractor employees performing services under this order will maintain the patient’s privacy act under the “HIPAA” laws. Contract employee performing services will be controlled, directed and supervised at all times by management personnel of the contractor. The contractor’s management shall ensure that employees properly comply with the performance standards outlined in the Statement of Work. Contractor employees will perform independent of and without the supervision of any Government official. Actions of contractor employees may not be interpreted or implemented in any manner that results in any contractor employee creating or modifying Federal policy, obligating the appropriated funds of the U.S. Government, overseeing the work of Federal employees, providing direct personal services to any Federal employee or otherwise violating the prohibitions set forth in Parts 7.5 and 37.1 of the Federal Acquisition Regulations (FAR). The Government will control access to the facility and will perform the inspection and acceptance of the completed work.

INVOICE REQUIREMENTS

1) One invoice per year (itemized) for all products provided

2) Total for entire invoice

A. The contractor shall submit the original and a copy of the invoices concurrently as follows:

a. An original to the following designated payment office:

National Institutes of Health

Office of Financial Management

Commercial Accounts, Room 4B-432

2115 East Jefferson Street MSC 8500

Bethesda, Maryland 20892-8500

b. One copy to the Administrative Officer at:

National Institutes of Health

Critical Care Medicine Department, Clinical Center

9000 Rockville Pike

Building 10, Room 2C145

Bethesda, Maryland 20892

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