Statement of Work.pdf

FEDERAL AVIATION ADMINISTRATION

CIVIL AEROSPACE MEDICAL INSTITUTE

FUNCTIONAL GENOMICS RESEARCH

LABORATORY

Sleep Deprivation Study for Aviation Research

“COMPARISON ACROSS MULTIPLE TYPES OF SLEEP DEPRIVATION”

STATEMENT OF WORK

Sleep Deprivation Study for Aviation Research

“COMPARISON ACROSS MULTIPLE TYPES OF SLEEP DEPRIVATION”

REVISED JULY 1, 2019

STATEMENT OF WORK

1.0 Introduction and Overview

The Civil Aerospace Medical Institute (CAMI) is the medical certification, education, research, and occupational medicine wing of the Office of Aerospace Medicine (AAM) under the auspices of the Federal Aviation Administration (FAA) Office of Aviation Safety (AVS). The mission of the Aerospace Medical Research Division of CAMI, located at the Mike Monroney Aeronautical Center (MMAC), in Oklahoma City, OK, is “to develop new and innovative ways to support FAA regulatory and advisory missions to improve the safety of humans in civilian aerospace operations.” As such, its goal is to “Continuously maintain the Absorptive Capacity for applying Science, Medicine, Bioengineering, and Technology to unrelentingly enhance human safety.”

The aerospace medical research program has been formulated to keep abreast of emerging human safety risk issues such as those brought by (1) the aging pilot population and changes in their health condition;

(2) advances in pharmacology, therapeutic tools, and surgical procedures, and (3) improved aircraft materials, equipment, cabin configurations, life support systems, and evacuation assistive devices– all of which may affect survival during an aircraft accident. The program has also been designed to address the complexity of software, technology, and systems integration practices as these continue to evolve.

For example, advances in computational biology, modeling and simulation, and tools to facilitate the integration of very large data sets containing disparate information will lead to improved knowledge management and decision-making processes in aerospace medicine.

The Aerospace Medical Research Division’s Bioaeronautical Sciences Research Branch contains the Biomedical Research Section, a subset of which is the Functional Genomics research team. This team is a pioneer in biomarker research pertinent to aviation safety, and is the requiring organization.

1.1 Scope of Work

The purpose of this document is to procure a sleep research laboratory source for neurobehavioral data and biological sample acquisition, as well as personnel expertise, for sleep impairment biomarker identification and understanding in accordance with the requirements specified in this Statement of Work and associated documents.

1.2 Period of Performance

Basic Year: September 18, 2019 through September 17, 2020 Option Year 1: September 18, 2020 through September 17, 2021 Option Year 2: September 18, 2021 through September 17, 2022 Option Year 3: September 18, 2022 through September 17, 2023 Option Year 4: September 18, 2023 through September 17, 2024

1.3 Requirement

1.3.1 Overview

The Federal Aviation Administration (FAA) Civil Aerospace Medical Institute (CAMI) is interested in discovering biomarkers associated with neurobehavioral or cognitive impairment during sleep loss and mistimed sleep. To achieve this goal, the Functional Genomics research team at CAMI has a requirement for blood samples to analyze gene expression profiles in individuals exposed to total sleep deprivation, sleep restriction, mistimed and restricted sleep simulating the schedule of a nightshift worker, and a well-rested control condition. Additionally, the team needs to acquire neurobehavioral data. Finally, cheek swab samples are needed to assay Single Nucleotide Polymorphisms (SNPs) or similar genetic variants pertinent to sleep, wakefulness, circadian rhythms, and/or neurobehavioral performance. The team will use this information to assess gene expression changes associated with reduced and mistimed sleep, to explore the relationship between gene expression and neurobehavioral changes from reduced and mistimed sleep, to enhance general understanding of changes during reduced and mistimed sleep, and to analyze the concordance of sleep and circadian rhythm-related genetic variants with this study’s results. Such genetic variants may help inform interpretation of the study’s findings, for example, by improving knowledge of the extent of genetic diversity and sleep-pertinent polymorphisms represented in the study population.

Details of the study not specifically addressed in this SOW and associated documents, including exhibits and contract clauses, are to be developed by mutual agreement of the Functional Genomics team and Contractor, subject to final approval by the Functional Genomics team. The Contractor must collect all samples (blood, cheek swab) and data (including neurobehavioral results) to address the functions specified herein, on each and every study subject.

The Functional Genomics team requires the Contractor to follow the sample collection schedule specified in Exhibit A - Table 1, to include blood collections at 4 h intervals across time awake as specified in four subject groups: Total Sleep Deprivation (TSD); Chronic Sleep Restriction – Night Sleep (CSR-N); Chronic Sleep Restriction – Day Sleep (CSR-D); and Control (CTL). The Functional Genomics team anticipates comparing results of each sleep loss group (CSR-D, CSR-N, TSD) to the CTL, as well as conducting analyses across all four conditions, including assessment of neurobehavioral data towards identifying biomarkers associated with performance impairment. Some blood draws are omitted for CSR-D and CSR-N subjects during experimental days to minimize the burden on study subjects and researchers (Exhibit A - Table 1).

1.3.2 Study subjects

The Contractor must deliver biological samples and neurobehavioral data from a total of 80 subjects.

Successful blood draws yielding blood of an acceptable quality level must be collected by the Contractor from a total of 20 subjects for each of the CTL, TSD, CSR-D, and CSR-N groups, with each subject following the schedule specified in Exhibit A - Table 1 for their group.

Each study subject must only participate in one of the four study condition groups; subjects cannot participate in more than one study condition group. Each study subject must not be a family member or close relative of any other study subject. For this purpose family members include close biological relatives and those living in the same household (e.g., spouse/partner or roommate). Although it is understood that respect for subjects’ privacy precludes some options for preventing recruitment of family members, reasonable efforts must be made. These should include at minimum measures such as requiring potential participants to attest that no other members of their family or household are participating in the study to the best of their knowledge, and only allowing one occurrence of a last name among study subjects if it may be reasonably anticipated that potential participants sharing that last name belong to the same family.

All subjects must be healthy adults (age 22-50, BMI 18.5-29.9, not obese) with no prior history of sleep or circadian disorders. Subjects must not represent the extremes of early-morning lark/night owl, and must have a habitual pattern of nightly sleep representative of a typical healthy adult with an adequate/normal sleep schedule. The presence of normal sleep patterns in each subject, to include absence of sleep disorders such as sleep apnea or restless leg syndrome, must be ascertained by the Contractor during acclimation/baseline days and/or during the recruitment process, using standard industry practice to include actigraphy data and polysomnography data collected by the Contractor (see section on Sleep Monitoring). Subjects must not use stimulants or depressants, alcohol, tobacco, medications including over-the-counter melatonin, or caffeine for a minimum of 1 week prior to arrival to the sleep laboratory and continuing through departure (Exhibit A - Table 1). Allowance of subjects’ use of contraceptives as an exception to the exclusion of medications is to be negotiated between the Contractor and Functional Genomics team, subject to the Functional Genomics team’s approval.

Ethnicity of the subject population must be representative of the local metropolitan area at the Contractor’s facility, or representative of the national United States population. Subjects must include males and females in approximately even numbers (45-55% female in each of the four condition groups, based on biological sex). Key demographics (including age, biological sex, ethnicity) must be represented across the four condition groups in an approximately even distribution, with no more than 15% difference in relative proportions of key demographics between the 4 study condition groups.

Demographics must be sufficiently similar across the four condition groups that differences among or between groups cannot be reasonably attributed to differences in demographics.

Contractor plans for ensuring demographic information and subject selection/screening procedures meets the requirements and intent of this SOW must be approved by the Functional Genomics team prior to the start of subject recruitment. Results of subject screening/recruitment and associated data must be provided to the Functional Genomics team, with provision of demographic data for each study subject linked to their identity by use of de-identified subject codes to protect subjects’ privacy.

Demographic information provided to the Functional Genomics team must include the age, biological sex, height, ethnicity, and body weight of each study subject.

The Contractor must plan for potential attrition in each of the 4 study groups (e.g., recruitment and assays of additional subjects as necessary) to ensure that successful high-quality blood draws are obtained from 20 TSD, 20 CSR-N, 20 CSR-D, and 20 CTL subjects in accordance with Exhibit A - Table 1. The Contractor must be responsible for recruiting and retaining additional subjects as necessary in the event of attrition, i.e., the Contractor must collect samples and data in accordance with Exhibit A - Table 1 from additional subjects to replace subjects that did not complete the study.

The Contractor must ensure that each and every study subject meets these and all requirements specified in this Statement of Work and associated documents.

All aspects of subject screening, recruitment, and retention are the responsibility of the Contractor and must be done by the Contractor, including protocol development and payment of subjects as appropriate and necessary. Subjects must be randomly assigned to the four condition groups (TSD, CSR-D, CSR-N, and CTL); if the Contractor desires any assignment other than random they must first obtain written permission from the Functional Genomics team. Assignment must be done in a way to minimize batch effects or uncontrolled bias. Subject groups must be run through the experiment in an approximately random or stratified random design, negotiated with the Functional Genomics team and subject to final approval by the Functional Genomics team. That is, the Contractor CANNOT run all 20 subjects of one of the four condition groups through the study, followed by all subjects of the next group, and so on. However within each run of a small subset of subjects housed/assayed simultaneously, those subjects housed/assayed simultaneously may all undergo the same study group condition as long as proper precautions are taken to minimize batch effects and bias. For this purpose, a small subset is defined as no more than 20% of the subjects in the study condition group. If subjects are assayed at multiple (more than one) Contractor facilities, this must follow a random or stratified design with ideally even numbers of subjects but at minimum 35% of the subjects in each of the four condition groups assayed at each Contractor facility. Again proper precautions must be taken by the Contractor to minimize batch effects and bias.

The Contractor must maintain responsibility for sample and data security, including proper protections for personally identifiable information and protected health information collected and maintained by the Contractor. The Contractor must be responsible for subject safety, including the protection of the rights and welfare of human subjects. The Contractor must produce de-identified subject codes, and maintain the master key associating de-identified subject codes with the individual subject. This key and all subject records with privacy information must be maintained by the Contractor in a manner compliant with local and federal regulations and code such as 45 CFR part 46 particularly the Common Rule (https://www.hhs.gov/ohrp/regulations-and-policy/regulations/common-rule/index.html;

https://www.hhs.gov/ohrp/regulations-and-policy/regulations/45-cfr-46/index.html) and the DOT duplication 49 CFR 11, HIPAA (https://www.hhs.gov/hipaa/index.html) as applicable, and IRB requirements, including regulations for retention and disposal of such information. The Contractor must comply with Government requirements for records retention. The work done by the Contractor must be in compliance with 49 CFR Part 11 (Protection of Human Subjects) as interpreted by the FAA and its Institutional Review Board.

1.3.3 Housing of subjects and facilities

The Contractor must house subjects in the Contractor’s facility for their entire time in the study (arrival through departure as specified in Exhibit A - Table 1). Bedding and mattresses must be maintained at an acceptable age and quality conducive to sufficient and consistent sleep quality among participants, and housing facilities must be well maintained and remain in good condition. The Contractor must ensure that housing for all subjects will be equivalent (to include lighting of <100 lux during time awake, lights off during scheduled sleep times, surveillance, access to non-vigorous activities, etc.)

such that differences between subjects, across days/times, and across study condition groups could not be reasonably attributed to differences in housing experience including facilities. Exposure to lighting from electronic devices, communication between subjects and individuals outside the sleep laboratory (e.g., calls/texts with friends and family), and ability to view real-time events outside the laboratory (e.g., television/newspaper/smartphone/internet access) will be negotiated and jointly decided by the Contractor and Functional Genomics team. However, the Functional Genomics team will have the final decision authority on allowed exposures. Exposures must be approximately even for subjects across the four condition groups; specifically the time and type of exposure allowed during time in the laboratory must be equivalent for all study subjects.

The Contractor must provide a weight-maintenance metabolic diet to all study subjects while they are in the sleep laboratory, and must obtain a self-report diet diary from subjects for the week prior to their entry into the sleep laboratory. The Contractor must provide the Functional Genomics team information on the diet provided to subjects while in the sleep laboratory, including description of food and beverage (besides water) items provided to and eaten by subjects with caloric value. Level of detail in provision of information about consumed food will be mutually negotiated between the Contractor and Functional Genomics team, with the understanding that consumption levels may be approximate. Levels of melatonin in food must be approximately consistent across meals and across all https://www.hhs.gov/ohrp/regulations-and-policy/regulations/45-cfr-46/index.html subjects during their time in the sleep laboratory, such that diet while in the sleep laboratory cannot be reasonably expected to lead to differences in circadian rhythm between subjects or across timepoints.

While in the sleep laboratory, subjects will be under video surveillance by the Contractor, and when necessary/appropriate have interaction with the Contractor’s staff to ensure adherence to the study protocol, particularly that subjects do not sleep during scheduled wake times. There must be video surveillance in the bedrooms, and the Contractor must either have video surveillance or an equivalent protocol (e.g., direct monitoring) to ensure subjects are awake in any common areas and adhere to the study schedule. Items for subject amusement involving non-vigorous activity, such as board games, must be available to subjects during intervals when no activity is scheduled (empty cells in Exhibit A - Table 1, i.e., times when sleep, neurobehavioral testing, blood draws, or meals are not scheduled).

Subjects must not undergo vigorous activity during their time in the sleep laboratory.

The Contractor must have -20°C freezers as well as -70°C or -80°C freezers, and adequate space in freezers, for storing blood samples until they can be shipped to and received by the Functional Genomics team at an address to be provided by the Functional Genomics team after Contract award. Scheduling of sample shipments must be coordinated with the Functional Genomics team to ensure their receipt. The sleep laboratory must have controlled lighting (fixed at <100 lux during time awake, lights off during sleep) and temperature (~21°C ±1) in all areas occupied by subjects, with minimal noise so as to not disturb subjects during time in bed. Each subject must have his/her own private room and bed, and adequate access to restroom/shower facilities.

Subjects must NOT undergo a constant routine protocol (i.e., subjects must not be forced to maintain a semi-recumbent position during their entire time in the sleep laboratory). The Contractor must provide individual bedrooms for each subject, adequate access to restroom/shower facilities with reasonable accommodation for privacy, and an adequate cleaning and maintenance regimen.

1.3.4. Sleep monitoring

To allow profiling of sleep, a total of three nights of polysomnography data must be collected per subject, for all subjects in each of the 4 study condition groups. Polysomnography data must be collected by the Contractor once overnight after the second acclimation day and preceding the baseline day for sleep loss treatment subjects (CSR-N, CSR-D, TSD) and the corresponding night for the CTL subjects. Additionally, polysomnography data must be collected by the Contractor during the two recovery nights of sleep for subjects in all 4 study condition groups. (For TSD, polysomnography is during sleep on the two nights of 10 h sleep following the experimental phase of total sleep deprivation, as on Exhibit A – Table 1). The Contractor must provide the Functional Genomics team with both raw and interpreted polysomnography data. The Contractor must ensure reasonable efforts are made to minimize bias from differences in subjective interpretations among different persons who score/interpret polysomnography results, if more than one person performs this task.

Subjects must be monitored by the Contractor for a minimum of one week prior to the sleep study to ensure conformance to a regular sleep schedule representing a well-rested condition of nighttime sleep, and refraining from naps. This monitoring must include subjects’ self-report sleep diary or equivalent, as well as actigraphy. This data including actigraphy must continue to be collected for the duration of the study (arrival through departure as in Exhibit A – Table 1, inclusive) to monitor time slept.

All data including polysomnography, sleep diary or equivalent, actigraphy, and dietary information must be provided by the Contractor to the Functional Genomics team, to include raw data and transformation of any raw data as appropriate into tabular format ready for review and statistical analysis.

1.3.5. Study schedule

Subjects in all four study condition groups will have slightly over one day of acclimation time in the sleep laboratory upon arrival to include time for training on neurobehavioral tests by the Contractor as appropriate; see Exhibit A - Table 1. Except meals and sleep times, activities are not specified during acclimation days, but subjects must be trained by the Contractor on neurobehavioral tests as appropriate during the acclimation stage without altering sleep or meal times from what is specified in Exhibit A - Table 1. This training must include sufficient repetition of tests to smooth out the learning curve during acclimation time, such that the impact of learning on neurobehavioral test results during baseline through recovery/post-recovery days is minimized.

Blood draws for gene expression along with neurobehavioral data collection will be performed during one baseline day after the acclimation sleep and immediately prior to the experimental phase; during the experimental phase during which sleep reduction occurs (TSD, CSR-N, CSR-D) or the well-rested condition is maintained (CTL group); and during recovery after the experimental phase (Exhibit A - Table 1). For TSD, blood draws also will occur during post-recovery days as indicated in Exhibit A – Table 1. At each sampling timepoint during time awake, subjects must first perform the neurobehavioral test battery, second undergo the blood draw, and third receive a meal as specified in Exhibit A - Table 1.

For acclimation/baseline and recovery nights, subjects in the CSR and TSD groups will receive a well-rested condition of 10 h time in bed (sleep times of 20:45-06:45 h), as will CTL subjects during the acclimation nights (first two nights in the laboratory prior to the baseline day; see Exhibit A - Table 1).

For the remainder of their time in the study when CTL subjects are assumed to be sleep satiated, CTL subjects will receive 8 h time in bed (22:45-06:45 h). During the experimental phases the TSD group will not sleep and instead will undergo 62 h continued wakefulness, while the CSR-D will receive 5 hours of sleep each day (09:45-14:45 h) and the CSR-N will receive 5 hours of nighttime sleep (21:45- 02:45 h) as specified in Exhibit A - Table 1. For the TSD, there will be a post-recovery phase to extend their duration in the sleep laboratory to be the same number of days as that of the other study condition groups, with required activities as designated in Exhibit A – Table 1. During all study phases, lights must be turned off during scheduled sleep times, and reasonable effort should be made to promote subjects’ sleeping during their scheduled sleep times.

1.3.6. Neurobehavioral data

Performance on a number of neurobehavioral assays must be measured by the Contractor at 4 hour intervals during waking periods in all groups, with timing of these assays to be right before blood draws (Exhibit A - Table 1). Each run of the neurobehavioral test battery must be completed by each subject in 45 min. or less to minimize performance decline due to boredom with the test battery. Order of individual tests within the battery at the within- and between-subject levels will be mutually negotiated between the Contractor and Functional Genomics team, with the intent of minimizing bias in results. The Contractor must provide staffing as appropriate to motivate subjects in performing the tests, inasmuch as possible promoting equal, consistent, and high motivation across subjects and timepoints to minimize noise in the data from motivation declines. The Contractor must be instrumental in monitoring results, including quality control and assessment of neurobehavioral test administration and findings. Specific neurobehavioral tests (objective and subjective) must be proposed by the Contractor. However, the objective neurobehavioral tests must include the 10 min.

Psychomotor Vigilance Test, PVT (https://link.springer.com/article/10.3758/BF03200977;

https://link.springer.com/article/10.3758/BF03200977 https://www.taylorfrancis.com/books/e/9780203998007/chapters/10.3109%2F9780203998007-10 ), with data for response time and number of lapses. Each and every study subject must undergo the same neurobehavioral tests. Neurobehavioral objective assays must encompass a series of tests to capture elements of all of the following:

• sustained attention/vigilance

• decision-making

• response to feedback stimuli

• spatial orientation

• memory/recall

• problem solving

• coordination

• inhibition/risk-taking behavior

• executive functioning/reasoning

• processing speed/ability to think and respond quickly

Additionally, the 45-min. assay period must include 2 surveys to capture the following subjective elements of neurobehavioral performance:

• self-diagnosed sleepiness levels (using a well-known method such as the Karolinska sleepiness scale (https://www.ncbi.nlm.nih.gov/pubmed/2265922)

• mood, particularly as applicable to identifying motivation declines for the test battery

The Contractor must use neurobehavioral tests and surveys that are well-validated through multiple publications and accepted by the general sleep research community, to enhance interpretation and general understanding of results. When possible the Contractor must use quantitative rather than qualitative assays. The multiple sleep latency test (or equivalent) must not be performed, to avoid the possibility of subjects falling asleep. When possible, the Contractor must choose neurobehavioral tests that minimize test-learning and introduction of testing bias from repeated test administration.

It is desirable to not confound gene expression or neurobehavioral results with typical post-waking sleep inertia. Therefore, the Contractor must ensure that subjects have been awake for at least 30 min.

prior to beginning neurobehavioral data collection. Although the objective and subjective neurobehavioral tests including surveys will be proposed by the Contractor and will be jointly selected by the Contractor and Functional Genomics team, the Functional Genomics research team will have the final decision authority for approval (acceptance, or rejection and requirement of an alternate test/survey).

Failure rate in neurobehavioral test data collection must not exceed more than 3 timepoints per study subject, and no more than 2 timepoints with a failed neurobehavioral test data collection may be consecutive. Within each study condition group (CTL, TSD, CSR-D, CSR-N) no more than 10% of the study subjects may have a failed neurobehavioral test. Failures will include refusal or inability of the study subject to complete the test, and/or instrument malfunctions or other errors that result in data being omitted, not collected, or unreliable.

1.3.7. Blood samples

Altogether a total of 27 blood samples for RNA-based gene expression work will be collected per TSD subject, 26 per CSR-N subject, 26 per CSR-D subject, and 31 per CTL subject. With 20 subjects in each group, this yields a grand total of 2200 successful high-quality blood samples, correctly collected, for gene expression (Exhibit A - Table 1). Each of these blood samples will have associated neurobehavioral test data collected immediately prior to the blood draw (Exhibit A - Table 1). Failure https://www.taylorfrancis.com/books/e/9780203998007/chapters/10.3109%2F9780203998007-10 rate in blood collection (inability to obtain and provide to the Functional Genomics team successful high-quality blood draws) for any given subject must not be higher than 3 blood draws per subject, and no more than 40% of the subjects in each of the four study condition groups may have a failed blood draw. Failures must be approximately random across timepoints (e.g., it cannot be that 40% of subjects failed to have the first blood draw collected). In general, RNA extracted from successful high-quality blood draws collected in PaxGene® RNA tubes, and processed according to the manufacturer’s protocol, yields high quality intact RNA when extracted via automated methods such as those typically used by the Functional Genomics team. Blood draws by the Contractor that do not yield acceptable quality level RNA may be considered failures (i.e. not counted as a successful high-quality blood draw), if both the Contractor and the Functional Genomics team mutually agree that the low quality results can reasonably be attributed to improper specimen handling, storage, and/or shipment by the Contractor.

No blood samples will be taken during the time in bed for scheduled sleep, unless mutually agreed to by the Contractor and Functional Genomics team.

Blood samples must be handled by the Contractor according to protocols provided by the Functional Genomics team. Blood samples must be taken from intravenous lines inserted at appropriate times by the Contractor, with two exceptions. One exception is that for post-recovery days, the Contractor may propose an alternate (e.g., needle) draw. A second exception is use of alternate protocols for blood draws to allow blood collection if intravenous lines fail at a limited number of timepoints (e.g., butterfly needle draw if the intravenous line clogs). These exceptions if desired by the Contractor, must be proposed by the Contractor during the stage of IRB protocol development and must be approved by the Functional Genomics team and IRB. If the Functional Genomics research team has not approved the alternate blood draw protocol prior to finalization of the IRB (see 4.1), then all blood draws must be collected from intravenous lines. At 4 hour intervals, specifically at times listed in Exhibit A - Table 1, blood will be drawn consisting of an aliquot of (~2.5 mL/tube, plus a few mL to clear the line before aliquot collection as appropriate) to be collected in a PAXgene® tube for gene expression. The line must be cleared PRIOR to collection of the blood into the PAXgene® tube. The Contractor must have a trained phlebotomist or nurse (as appropriate and required by local policy and regulations applicable at the Contractor’s facility, and the IRB) perform intravenous line insertion and removal, and perform blood draws. The Contractor will be responsible for proper handling of blood and other potentially infectious material, and for proper handling and disposal of biohazards as appropriate.

The FAA Functional Genomics team will provide protocols for preservation of samples (e.g., protocol for inverting, incubating, and freezing PAXgene® tubes) that must be followed by the Contractor. The Contractor will be responsible for proposing (subject to the Functional Genomics team approval) and following the phlebotomy protocol of drawing blood including reinsertion of lines when needed, clearing of lines, collection of blood into PAXgene® tubes, removal of lines, and associated tasks. As part of handling blood samples, the Contractor must add the appropriate volume of blood to the PAXgene® tube, neither over nor under-filling, and must follow instructions for appropriate mixing/inversion, incubation, freezing, storage and shipment of blood samples. The Contractor must be responsible for subject safety as well as for the safety and training of the Contractor’s personnel, including bloodborne pathogen training and ethical conduct in human research (e.g., Collaborative Institutional Training Initiative, https://about.citiprogram.org/en/homepage/) and certification of personnel, as appropriate. The Contractor must provide all blood samples to the Functional Genomics team. Initial protocol testing and practice of the blood draws by the Contractor will be coordinated as mutually agreed by the Functional Genomics team and Contractor.

The Contractor is expected to make reasonable efforts to ensure study subjects maintain sufficient hydration levels to facilitate collection of blood draws.

1.3.8 Cheek Swab Samples

Two cheek swabs per subject must be collected by the Contractor for DNA, to allow assessment of SNPs or other genetic variants relevant to sleep, wake, circadian rhythms, and/or neurobehavioral performance. The DNA samples can be collected on any day(s), at any time during time awake that does not interfere with collection of the other samples or data, and that does not interfere with any other scheduled activity (Exhibit A - Table 1). The Contractor must provide all completed swabs to the Functional Genomics team.

No more than 10% of study subjects in each study condition group may have a failure in one of the two cheek swab collections. The Contractor must provide at minimum one cheek swab per study subject to the Functional Genomics team; failure to provide at least one will be considered an event of attrition of the study subject.

1.3.9. Data Provision and Interpretation

The Contractor must be directly involved in analysis and interpretation of data and provision of this information to the Functional Genomics team, including objective and subjective neurobehavioral assay results, actigraphy data, and polysomnography data. The Contractor must provide text (in a mutually agreed format) regarding study methods in the sleep laboratory, including all details relevant for data interpretation and publication. The Contractor must provide all data gathered to the Functional Genomics team, including all neurobehavioral data and de-identified subject demographics (e.g., age, biological sex) so the Functional Genomics team may test for relationships of this data with gene expression and genetic variants, and computationally address batch effects as appropriate. Legends, descriptions, and interpretations of data and study methods must be provided by the Contractor, to include clarification of any abbreviations used and explanations as requested by the Functional Genomics team. The Contractor must provide all raw data to the Functional Genomics Team, as well as conduct any initial data processing or scoring required to render data (including surveys and other neurobehavioral test results, and actigraphy data) in tabular format ready for statistical analysis, and provide both raw and processed polysomnographic results. Polysomnography processing must be done by a certified polysomnographist. See contract clauses and Section H – Special Contract Requirements of the Terms and Conditions, for details on data ownership.

In short, all data germane to analysis, publication, and data archival must be provided to the Functional Genomics team (see Section H – Special Contract Requirements of the Terms and Conditions). Any issues that arise with sample or data collection or the study subjects, or notes regarding issues that might reasonably be anticipated to have an impact on the sample/data or their interpretation, must be described to the Functional Genomics team. This description must be provided as part of the requested monthly updates described in SOW section 4.3.

A single set of de-identified subject codes must be used consistently across all collected data and biological samples, and in the event the study is conducted in multiple locations (more than one facility), the subject coding system must be coordinated between all facilities such that each subject identifier is unique to an individual participant. The Contractor will be responsible for ensuring labelling is clear and legible to the Functional Genomics team. The specific type of labeling (e.g,. use of a barcode labeling system) will be selected by negotiation between the Functional Genomics team and Contractor. The Contractor will be responsible for maintaining subject information including contact information as appropriate, in accordance with requirements of the IRB and any applicable policies, laws, or regulations (including 49 CFR 11, and including HIPAA and 45 CFR 46 as appropriate), with appropriate protection for Privacy Health Information and Personally Identifying Information.

Provision of results must include information and labels required for associating all data and all biological samples with the following: the time of collection, calendar date, day in the sleep laboratory as in Exhibit A - Table 1 (or number of days prior to arrival, for items collected before arrival), condition group (TSD, CSR-D, CSR-N, or CTL), and individual subject as represented by a unique de-identified subject code. In any case where either sample (e.g., blood tube or cheek swab) or data is incorrectly labelled and the label is not corrected by the Contractor, including cases of provision of samples entirely missing a label or where the sample/data cannot be associated with the information requested in this SOW, the sample or data will be considered a failure. The sample or data must still be offered to the Functional Genomics team and provided if requested by the Functional Genomics team, but it will not be considered a successful collection (e.g., a blood tube without a label would not be counted as a successful high-quality blood draw). The subject code must allow association of the individual with their pertinent demographic data, and facility if multiple (more than one) Contractor facilities are used to house subjects.

1.4.0 Institutional Review Board

The Contractor must develop a study protocol for Institutional Review Board (IRB) approval, and coordinate with the Functional Genomics Team in the development and review of this protocol. The FAA will determine whether the IRB approval must be obtained by the institution of the Contractor and/or the FAA. The Contractor must ensure the protocol meets the requirements specified in this Statement of Work and associated documents. The Contractor must work with CAMI to ensure all documentation, including IRB approval, subject written informed consent, and other paperwork, is completed in a timely fashion. The Contractor will notify the Functional Genomics Team within five

(5) business days in the event of an unscheduled change in sample collection. The Contractor must be responsible for IRB approval and keeping the IRB agreement active throughout the duration of the study as appropriate (see Deliverables 4.1). In all cases keeping an IRB agreement active includes conducting amendments and extensions as needed. If the FAA determines that IRB review will take place by the FAA, the Contractor must provide support as requested by FAA personnel for IRB review, including completing requested documentation and responses to requests for information.

The Contractor must follow the IRB-approved protocol, and will be responsible for obtaining and maintaining written informed consent of all subjects. Use of broad consent (https://www.hhs.gov/ohrp/sachrp-committee/recommendations/attachment-c-august-2- 2017/index.html) must be done to the extent possible and permissible by IRB requirements, to permit future use of collected samples and data. Selection of consent type and its contents must be done in compliance with IRB requirements.

The Contractor must obtain from each and every study subject written informed consent for not only the work and collection of samples and data in this study, but also for archival of the data and results of this study. Consent for archival is to include consent for archival in a public repository (e.g., dbGaP, Dryad Digital Repository) of neurobehavioral performance data, genetic data including sequences, genetic variant information, gene expression levels, study subject demographics, sleep diary or equivalent data, diet data, actigraphy and polysomnography data, and other results and information collected in this study.

If over and above options are pursued, the IRB must include consent for the additional work and archival of the data. The Contractor should expect the need for data archival consistent with publication in the peer-reviewed literature of findings from the results of this study, and must ensure study subjects are properly consented.

The Contractor must ensure consent of study subjects is compliant with 49 CFR 11 and must ensure the consent permits all research described in this SOW and associated documents, including that described in exhibits as appropriate. Consent must include processing of data and of the biological samples using approaches such as RNA-Sequencing or microarrays, as well as SNP and genetic variant analysis. The Contractor must maintain copies of the signed consent forms in accordance with the requirements of 49 CFR 11 (Protection of Human Subjects) as interpreted by the FAA and its Institutional Review Board, and the requirements of the IRB overseeing this study.

The Functional Genomics team reserves the right to send a representative to the Contractor’s facility for purposes such as periodic supervision and collaborative discussions, with a minimum of 2 weeks notice.

Reasonable accommodation will be made for the Contractor’s schedule, but it is expected that the Contractor will be responsive in a timely manner to questions or requests for collaborative discussions, including teleconferences and on-site visits by the Functional Genomics team. It is anticipated there will be a kick-off meeting post-award, one or more additional collaborative discussions during the IRB review process, and an annual collaborative discussion. Exact frequency and use of on-site vs.

teleconference venues will depend on the needs that arise during performance of work.

However the Contractor will be expected to carry out functions autonomously subject to the mutual agreements, negotiations, approvals, and other constraints as set forth in this SOW and associated documents, including contract clauses and Section H – Special Contract Requirements of the Terms and Conditions. The Contractor will be expected to have and provide expertise for interpretation of data, including neurobehavioral data, actigraphy, sleep and diet diaries, and polysomnography results.

The Contractor must designate a primary Point of Contact who must be responsible for responding to questions and concerns of the Functional Genomics team and other FAA personnel as appropriate, and provide the name and contact information for this individual. In the event the primary Point of Contact is out of the office or otherwise unavailable, the Contractor must designate in advance an alternate Point of Contact.

1.4.1 Travel

The government shall reimburse the Contractor for travel costs incurred only in the performance of this contract. Any travel supported by this contract must be coordinated with the COR and be in accordance with Federal Travel Regulations.

2.0 Place of Performance:

Subject screening and recruitment, subject housing, exposure of individuals to the appropriate conditions (TSD, CSR-D, CSR-N, CTL), neurobehavioral assays including surveys, collection of samples, handling and preservation of samples until shipment to the Functional Genomics team, sample shipment, and all activities in Exhibit A - Table 1 will be performed by the Contractor at the Contractor’s facility. In short, all work performed by the Contractor will be done at the Contractor’s facility.

2.1 Government Furnished Materials and Services Support

The Government intends to provide the following materials via direct shipment of the items or payment to the Contractor for purchase by Contractor: PAXgene® Blood RNA tubes for sample collection, collection materials for harvesting DNA cheek swabs, and protocols for handling and preserving collected blood prior to shipment to the Functional Genomics team at CAMI. The Functional Genomics Team will also provide protocols (e.g., overnight shipment on dry ice) for shipment of biological samples, and the address to which samples must be shipped. Protocols provided by the Functional Genomics team must be followed by the Contractor; any deviations must be approved in advance in writing by the Functional Genomics team. The Government will provide the Contractor a mutually-agreed upon number of PAXgene® Blood RNA tubes for initial protocol testing, if the need for such testing is mutually agreed upon by the Functional Genomics team and Contractor. Timing and method for provision of materials by the Government will be via mutual agreement of the Functional Genomics team and Contractor.

2.2 Contractor Responsibility

The Contractor must provide all facilities, equipment, personnel, training, and materials not explicitly listed in section 2.1 above of Government Furnished Services, including all items required for neurobehavioral data collection. The Contractor must ship all blood samples via priority overnight delivery on dry ice, with samples shipped in Styrofoam boxes (or functional equivalent to Styrofoam for keeping samples cold and protected), and must ship all samples including cheek swabs in accordance with shipping requirements to be provided by the Functional Genomics Team.

All aspects of recruiting, monitoring, and housing of study subjects must be the responsibility of the Contractor. The Contractor also must be responsible for subject safety and well-being including full responsibility for any adverse events and unanticipated problems involving risks to subjects or others.

The Contractor must be responsible for the safety and well-being of its staff/personnel. The Contractor must be responsible for the security and confidentiality of any subject data and records, compliance with 49 CFR 11 as interpreted by the FAA and its IRB and compliance with other regulations (e.g., HIPAA) if and where applicable, and notification of study subjects of findings or items associated with this study if such notification is required (e.g. in the IRB or via legal requirements).

The Contractor must be responsible for the IRB process and approval at their institution (if IRB review at their institution is required by the FAA), subsequent IRB extensions/amendments as appropriate, the informed consent process, and IRB compliance. The Contractor must be responsible for providing any paperwork and information required to obtain IRB approval by the FAA if the FAA elects to perform IRB review, including that required for subsequent IRB extensions/amendments as appropriate, and will maintain responsibility for the informed consent process. In short, the Contractor must be fully responsible for any IRB work and approval at their facility and must provide support as requested by the Functional Genomics research team or FAA personnel for IRB review by the FAA, subject to the FAA’s decision of where IRB review occurs. Regardless of where IRB review occurs, the Contractor must comply with requirements of the IRB and IRB protocols, including reporting requirements, and will assume primary responsibility for IRB approval.

The Contractor must be responsible for ensuring adherence to the study schedule (Exhibit A - Table 1), including ensuring that subjects remain awake during wake-designated times in the sleep laboratory and only sleep during the times designated for sleep. Experienced personnel for blood and neurobehavioral data collection must be provided by the Contractor including qualified phlebotomists/nurses as appropriate for intravenous line insertion, intravenous line removal, and blood draws; laboratory technicians, support staff, research assistants and supervisors as necessary; polysomnography experts;

and neurobehavioral experts. The Contractor must be responsible for ensuring personnel have the appropriate training and certification levels, including but not limited to bloodborne pathogen and ethical conduct in human subject research, as appropriate for the personnel and their roles. The Contractor must be responsible for all interaction with subjects, including provision of actigraphs to subjects prior to arrival in the sleep laboratory for monitoring as specified in the SOW section 1.3.4.

The Contractor also must provide intravenous lines, labels, video surveillance equipment, actigraphy equipment, polysomnography equipment and personnel for scoring data, equipment for conducting and personnel for scoring neurobehavioral tests, software as needed for data collection and conversion of data into tabular form ready for statistical analysis as appropriate, biohazard safe containers as needed, shipping expenses to include dry ice, and all other material costs associated with collecting, processing, storing, and shipping of samples, data, and deliverables not explicitly listed as a Government Furnished item above. The Contractor must have -20°C freezers, and -70° to -80°C freezers, with an appropriate backup power system and access to backup freezers in the event of freezer failure, and must be responsible for appropriate storage of biological samples until they can be shipped to and received by the Functional Genomics team. This includes appropriate storage of PAXgene® blood tubes, and storage of DNA cheek swabs.

The Contractor must provide the facility for conducting the sleep studies, which must be a sleep laboratory with controlled lighting (fixed at <100 lux during time awake, lights off during sleep), temperature (~21°C ±1), subject dietary and activity regimen. These limits on light exposure and temperature must exist for the private bedrooms and any other areas (e.g., restrooms, common areas) that the subjects may enter between their arrival and departure (Exhibit A - Table 1). The Contractor’s facility must include personnel for maintaining the sleep laboratory and subject supervision. The Contractor must ensure minimal noise levels, such as to not disturb subjects during time in bed for sleeping. Subjects must be housed and data collected on-site at the Contractor’s laboratory for the entire time they are in the study, arrival to departure as specified in Exhibit A - Table 1. The Contractor must provide each subject his/her own private room and bed, and adequate access to restroom/shower facilities in the sleep laboratory with reasonable accommodation for privacy. The Contractor must be responsible for maintaining and cleaning the facility used to house subjects.

If more than one Contractor facility is used, the Contractor must be responsible for coordination of protocols, housing, and subjects’ experiences across facilities to ensure differences among subjects and…