Menopausal Sleep Fragmentation: Impact on Body Fat Gain Biomarkers in Women
Phase: PHASE4
Conditions: Menopause
Sponsor: Brigham and Women's Hospital

Document Title: 
Study Protocol 
Protocol Title: 
Menopausal Sleep Fragmentation: Impact on Body Fat Gain Biomarkers in 
Women 
Protocol #: 
MGB Human Research Committee #2016P002821 
NCT:  
 
03047330 
Document date: 
January 3, 2024 

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Version Date: January 3, 2024 
Filename: Protocol Summary
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PRINCIPAL/OVERALL INVESTIGATOR 
Hadine Joffe, MD, MSc 
 
PROTOCOL TITLE 
Menopausal Sleep Fragmentation: Impact on Body Fat Gain Biomarkers in Women 
 
 
FUNDING 
National Institute of Health – National Institute on Aging 
 
 
VERSION DATE 
1/3/2024 
 
SPECIFIC AIMS 
 
 
Aim #1: To determine the impact of menopause-related sleep fragmentation on metabolic and 
behavioral biomarkers of body fat gain using an experimental sleep disruption paradigm to fragment 
sleep in healthy female volunteers 
Hypothesis 1a: Experimental sleep fragmentation will result in an adverse adipokine profile as a 
metabolic biomarker for body fat gain 
Hypothesis 1b: Experimental sleep fragmentation will result in increased hunger and caloric intake 
Hypothesis 1c: Sleep disruption due to hot flashes will result in an adverse adipokine profile and 
increased hunger and caloric intake 
Aim #2: To examine if hot flashes are associated with metabolic and behavioral biomarkers of body 
fat gain using an experimental hormone suppression paradigm in healthy women 
Hypothesis 2: Hot flash frequency will correlate with a worse adipokine profile and increased 
hunger and caloric intake 
Aim #3: To determine if estradiol withdrawal adversely affects metabolic and behavioral 
biomarkers of body fat gain using an experimental hormone suppression paradigm in healthy 
women 
Hypothesis 3: Experimental withdrawal of estradiol will correlate with a worse adipokine profile 
and increased hunger and caloric intake 
Aims 1–3 Exploratory Hypotheses: 
1) Experimental sleep fragmentation mimicking menopause-related sleep disruption will reduce 
resting energy expenditure. 2) Baseline levels of total body fat and visceral adiposity will influence 
PARTNERS HUMAN RESEARCH COMMITTEE 
PROTOCOL SUMMARY 
Answer all questions accurately and completely in order to provide the PHRC with the relevant 
information to assess the risk-benefit ratio for the study.  Do not leave sections blank. 
Concisely state the objectives of the study and the hypothesis being tested. 

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the metabolic and behavioral response to sleep fragmentation, hot flashes, and estradiol withdrawal. 
3) Negative mood state will correlate independently with an adverse adipokine profile and increased 
hunger and caloric intake. 
 
BACKGROUND AND SIGNIFICANCE 
 
Obesity is highly prevalent in midlife and older women. Over 50% of women gain body fat during 
the menopause transition, unrelated to chronological aging, increasing the likelihood of cardio-
metabolic diseases after menopause. While these body composition changes are attributed to E2 
withdrawal, other menopause-related factors likely contribute to adiposity because: 1) body fat 
gains are not uniform despite universal progression to hypo-estrogenism, and 2) body fat accrues 
during the MT, when E2 continues to be produced intermittently. Sleep fragmentation related to HF 
occurs in almost half of midlife women and may be an important factor driving risk for body fat 
gain, but its impact on metabolic and behavioral biomarkers of body fat gain has not been 
investigated. This sleep fragmentation results commonly from HF, which occur in 2/3 of midlife 
women. In other populations, individuals who sleep less are heavier and have adverse adipokine 
profiles. Experimental studies restricting TST show that sleep reduction lowers leptin, and increases 
ghrelin, appetitive drive, and caloric intake, key biomarkers of adiposity, with some sex differences 
in effects. However, TST is not reduced in HF-related sleep fragmentation, and metabolic effects 
of sleep fragmentation have received limited investigation in humans, although animal studies show 
that sleep fragmentation adversely effects adipokines and eating behaviors. The goal of the current 
proposal is to determine the impact of HF-related sleep fragmentation, as well as that of HF and E2 
withdrawal, on body fat gain in women. Given the prevalence of HF and associated sleep 
disruption, identifying the impact of these core menopause symptoms on body fat gain will inform 
health promotion strategies to prevent gains in body fat in midlife women, thereby reducing 
cardio-metabolic diseases in aging women. 
 
RESEARCH DESIGN AND METHODS 
 
Overview 
Forty women ages 18–45 years will complete a protocol involving open-label treatment with 
standard-dose depot leuprolide to induce hypo-E2 in all women and generate HF in approximately 
2/3, thereby mimicking menopause (see Schema). As part of the protocol, subjects will be admitted 
to the BWH Clinical And Translational Science Center/Center for Clinical Investigation (CCI) 
inpatient unit for a total of 10 nights over a 2-month period (see Schema). Assessments will be 
divided into 2 sleep blocks. Each SB includes 2 nights during which sleep is undisturbed and 3 
nights during which sleep is experimentally fragmented. Using PSG, sleep patterns will be 
experimentally interrupted to the same target level for all subjects during all fragmented nights using 
auditory procedures to mimic the menopausal hot flash-associated sleep maintenance problem. The 
in-lab sleep blocks will be conducted: 1) during the mid-to- late follicular phase of the menstrual 
Provide a brief paragraph summarizing prior experience important for understanding the 
proposed study and procedures. 
Briefly describe study design and anticipated enrollment, i.e., number of subjects to be enrolled 
by researchers study-wide and by Partners researchers.  Provide a brief summary of the 
eligibility criteria (for example, age range, gender, medical condition).  Include any local site 
restrictions, for example, “Enrollment at Partners will be limited to adults although the sponsor’s 
protocol is open to both children and adults.” 

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cycle when E2 is high; and 2) after at least 4 weeks on leuprolide when HF are prominent in a subset 
of subjects. 
 
Participants will be given a standardized eucaloric diet for several days preceding the inpatient 
admission, as well as for the first four days of the inpatient visit. On one day before admission, and 
on the final day of inpatient admissions, subjects will have an ad lib diet. Fasting adipokine levels 
will be measured and leptin levels will be obtained to determine the 24-hour leptin profile under 
standardized eucaloric diet conditions. Food intake and macronutrient composition will be assessed 
as key behavioral biomarkers for body fat gain when the diet is ad lib. Hunger will be measured 
under both eucaloric and ad lib dietary conditions. Secondary and exploratory measures of objective 
HF using skin conductance monitors, negative mood/affect, reward seeking behaviors, and 
appetite/food preference will also be obtained. Behavioral measures will continue to be assessed at 
home after the 2nd SB is complete. 
 
Protocol Schema 
 
 
 
 
 
Study Sample and Criteria 
Informed consent will be obtained from up to 130 healthy non-pregnant premenopausal women 18-
45 years who do not have hot flashes in order to have 40 women complete study procedures. 

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Informed consent will be obtained from up to 25 healthy non-pregnant premenopausal women 21-
45 years to complete the pilot study.  Eligibility will be evaluated initially through a telephone 
screening process and then established in an in-person screening office visit. The following 
inclusion and exclusion criteria will then be applied: 
 
A. Inclusion/Exclusion Criteria for Full Study Participants 
 
Inclusion Criteria: 
1. Healthy premenopausal women 18–45 years old 
2. Habitually sleep 7–9 hours per night and usual bedtime between 9 pm and 2 am 
3. Consume <2 alcoholic drinks per day 
4. Consume <3 caffeinated drinks per day 
5. Regular monthly menstrual cycles 
6. No lifetime history of hot flashes 
7. Willingness  to  use  barrier  methods  of  contraception  or  IUD during study and after 
completion of study until menses resume 
8. Weight >110 pounds (given blood sampling plan; see Section 1c below) 
9. Body mass index ≥18.5 and  <35 kg/m2 (to avoid possible under-dosing of leuprolide and 
eliminated confounding of adipokine effects by obesity) 
10. Good general health 
11. Ability to understand and willingness to sign written informed consent document 
 
Exclusion Criteria: 
1. Contraindication, hypersensitivity or previous adverse reaction to gonadotropin releasing 
hormone agonists 
2. Pregnancy, confirmed by serum HCG at screening and prior to leuprolide administration 
3. Breastfeeding 
4. Tobacco use 
5. Current or recent (1 month) use of centrally active medications, or a plan to initiate one of these 
medications during the study. 
6. Current or recent (1 month) use of systemic doses of female reproductive hormone medications, 
except from intrauterine device initiated more than 6 months prior to study participation 
7. Current or recent (1 month) use of over-the-counter (OTC) medications that may affect hot 
flashes, sleep, or mood (eg diphenhydramine, St. John’s Wort, Black Cohosh, and other 
phytoestrogen supplements) 
8. Shift workers or time zone travel within 4 weeks prior to beginning the study 
9. Obstructive  sleep  apnea  as  indicated  by  an  elevated  score  on  the  Berlin  screening 
questionnaire or by history 
10. Insomnia symptoms as indicated by a score on the Insomnia Severity Index suggesting 
subthreshold or clinically significant insomnia symptoms 
11. Contraindications to CT or DXA scanning 
12. Diagnosis of osteoporosis or osteopenia, or history of non-traumatic fractures 
13. Hypothalamic-pituitary-adrenal axis disorders 
14. Diabetes by history, or as indicated by an abnormal hemoglobin A1c 
15. Gastric bypass, metabolic disorders, or other conditions that affect adipokines 
16. Abnormalities on screening laboratory tests (hemoglobin A1c, complete blood count, thyroid, 

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hepatic and renal function) including specifically a hemoglobin <12 g/dl (given multiple blood 
draws) judged by the investigator to be medically relevant 
17. Hearing loss interfering with ability to respond to auditory stimuli 
18. Cardiovascular illness 
19. Neurological illness 
20. Psychiatric illness, defined by clinical assessment at baseline as: 
a. Current depression as indicated by an elevated score on the PHQ-8 
b. A lifetime history of major depressive disorder, bipolar disorder, eating disorder, or 
psychotic symptoms 
c. Current suicidal or homicidal ideation, or any prior suicide attempt or inpatient psychiatric 
hospitalization 
21. Current medical or psychiatric conditions or medicine use judged by the investigator to 
interfere with study procedures or data collection 
a. An alcohol or substance-use disorder active within the past year 
 
B. Inclusion/Exclusion Criteria for Pilot Study Participants 
 
Inclusion Criteria for Pilot Study Participants: 
1. Healthy premenopausal women 21–45 years old 
2. Habitually sleep 7–9 hours per night and usual bedtime before 12 midnight 
3. No lifetime history of hot flashes 
4. Body mass index <35 kg/m2 
5. Good general health 
6. Ability to understand and willingness to sign written informed consent document 
 
Exclusion Criteria: 
1. Current medical or psychiatric conditions or medicine use judged by the investigator to 
interfere with study procedures or data collection 
2. Pregnancy 
3. Breastfeeding 
4. Regular tobacco use 
5. Shift workers 
6. Diagnosis of obstructive sleep apnea 
7. Insomnia symptoms as indicated by a score on the Insomnia Severity Index suggesting 
subthreshold or clinically significant insomnia symptoms 
8. Hearing loss interfering with ability to respond to auditory stimuli 
9. Current depression, defined by a score >3 on the PHQ-2 
 
Off-Study Criteria: 
1. Emergence of significant psychiatric symptoms, suicidal or homicidal ideation 
2. Development of any significant medical problem 
3. Pregnancy 
4. Significant deviations from study protocol,  including enrollment  in  another  research 
protocol that may influence the results of the current protocol. 
 
 
 

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Study Procedures Overview 
 
 
Screening Phase (1-2 months) 
 Telephone/Pre-screening: Consent to screen will be obtained orally on the phone for the initial 
screening procedures and diaries. Women who verbally consent to screening and are interested in 
participating in this trial will answer some questions by phone to assess preliminary eligibility. The 
Berlin Questionnaire will be administered to screen for sleep apnea, and questions about general 
medical and mental health will be asked to screen for any major exclusion criteria. Those who are 
eligible and interested will be scheduled for an initial remote screening and research consent visit 
with a study clinician. 
  
 Screening visit 1a (remote): Screening visit 1a will be conducted remotely. Written informed 
consent will be obtained via electronic signature using REDCap e-consent template and initial 
eligibility criteria will be assessed by a clinician, including a medical and psychiatric interview, and 
sleep interview. All women who sign the consent form will be assigned a study identification 
number regardless of eligibility status. Eligible participants will start monitoring their menstrual 
cycles and tracking their sleep after this visit. 
 
 Screening Visit 1b (in-person): At this visit, a clinician will conduct a physical exam, and a blood 
draw will be taken for screening assays (e.g., CBC, glucose, insulin, HCG, TSH), for baseline levels 
of adipokines, metabolic and reproductive hormones, and for measures of immune function related 
to sleep and menopause. Once lab results are reviewed by a study clinician, final eligibility will be 
assessed. Eligible participants will be scheduled for Sleep Block 1, leuprolide injection, and dual-
energy x-ray absorptiometry (DXA) scan. 
 
Sleep Block 1 (10 days – two weeks before GnRH Agonist injection) 
 Subjects will be admitted for two five night stays within the BWH Clinical And Translational 
Science Center/Center for Clinical Investigation (CCI) inpatient unit. Subjects will also complete 
at-home procedures, including an at-home PSG procedure. Subjects may log meals before inpatient 
admission and for up to 7 days after discharge with the app MealLoggerTM 
 
GnRH Agonist Injection (1 day) 
 Subjects will come to one of the outpatient research units to be given an injection of leuprolide 
(GnRH agonist). The timing of ovulation will have been previously determined based on the 
collection of at least 3 menstrual cycle start dates. Leuprolide will be administered in the week prior 
to an expected menstrual cycle start date. If the subject starts her menstrual cycle early prior to 
receiving leuprolide, the protocol will be extended to the next cycle.On the day of the scheduled 
appointment for the injection, a urine pregnancy test will be performed to confirm that the subject 
is not pregnant and blood will be drawn to measure reproductive hormone levels. 
 
Sleep Block 2 (10 days – 4 weeks after GnRH Agonist Injection) 
 Subjects will be admitted for two five night stays within the BWH Clinical And Translational 
Briefly describe study procedures.  Include any local site restrictions, for example, “Subjects 
enrolled at Partners will not participate in the pharmacokinetic portion of the study.” Describe 
study endpoints. 

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Science Center/Center for Clinical Investigation (CCI) inpatient unit. Subjects will also complete 
at-home procedures, including an at-home PSG procedure. Subjects may log meals before inpatient 
admission and for up to 7 days after discharge with the app MealLoggerTM. 
 
Follow-up Phase (4-12 weeks or until symptoms resolve) 
 Follow-up phone call from the study coordinator 4 weeks after the final study visit and every 4 
weeks thereafter to confirm resumption of menses and cessation of hot flashes (if they occurred). 
 Study staff will also collect food and appetite measures that were recorded by subjects the end of 
Sleep Block 2 
 Subjects who do not have resumption of menses or who continue to have hot flashes 4 months after 
receiving the leuprolide injection (3 months after final visit) will be referred to their routine medical 
provider for evaluation. 
 A study summary letter will be sent to subjects once the final follow-up phone call has occurred. If 
there are any abnormal laboratory results listed in the letter that have not yet been discussed with 
the subject, a study clinician will call the subject to discuss prior to sending the letter. 
 
Detailed Description of Sleep Blocks 
Assessments will be divided into 2 sleep blocks, each of which lasts 9 nights/10 days and includes 
5 nights of inpatient laboratory assessment. Each block includes 2 in-lab nights during which sleep 
is undisturbed and 3 nights during which sleep is experimentally fragmented. Subjects will also 
receive an actigraphy watch and record their sleep habits before entering the lab, and for up to 7 
nights after. During one night of each sleep block, subjects will use an at-home PSG to record their 
sleep. Subjects will return to BWH the following morning to deposit the PSG device with study 
staff, and to eat an ad lib breakfast. 
 At-home procedures during the Sleep Block: Subjects will complete four days of at-home 
procedures before their admission to the CCI inpatient unit. On the 1st, 2nd, and 4th days, subjects 
will eat eucaloric meals provided by the BWH Nutrition Core. On the 3rd day, subjects will be given 
food provided by the Nutrition Core for that entire day, and will be allowed to consume as much as 
desired at home while logging their meals, snacks and drinks. On the 2nd night of the sleep block, 
subjects will come to Brigham and Women’s to have an at-home ambulatory PSG machine attached; 
this machine will be removed the following morning when subjects arrive for breakfast. Eucaloric 
diets will approximate the current American women’s diet, with approximately 15% protein, 50% 
carbohydrates, and 30% fat. Subjects will select each eucaloric meal from options prepared by the 
Nutrition Core; this selection will be done in advance of sleep blocks. If a subject would like to 
change their meal preferences, they will be permitted to do so as long as the change can be 
accommodated by the BWH Nutrition Core. They will be asked to finish meals and log any 
deviations of the eucaloric diet. For each day with an ad lib diet, caloric intake and macronutrient 
composition will be calculated and analyzed with Nutrition Data System for Research software after 
each meal and snack. Subjects will order these meals from a prepared ad lib menu. 
 Procedures to occur in both SBUD and SBSF: Subjects will remain indoors during each admission. 
Technicians will hook subjects up to a PSG monitor each night. Participants will be given a 
standardized eucaloric diet for the first 5 days of their visit, and will have an ad lib menu on the 6th 
day of their in-lab stay. On the 6th day, subjects will be given food provided by the Nutrition Core 
for that entire day, and will be allowed to consume as much as desired at home while logging their 
meals, snacks and drinks with the app MealLoggerTM. Subjects will be expected to finish these 
meals.  Caloric and macronutrient intake will be quantified. Fasting adipokine levels, other 

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metabolic and stress markers will be collected during each visit. Blood samples will be collected 
repeatedly up to every 30 minutes during each visit. Blood will be drawn without interrupting sleep 
using an approximately 72” long IV line from outside the room and a blood-sparing technique to 
minimize blood loss. If issues with the IV occur, then select samples, up to 2 per wake episode may 
be drawn via venipuncture. Core Body Temperature will be measured with an oral thermometer. 
Hunger will be measured under both eucaloric and ad lib dietary conditions by administering 
Hunger Visual Analog Scales. On the day of admission, reproductive hormone levels will be tested, 
a urine pregnancy test will be performed, and morphometric measures will be gathered.   During 
the inpatient stay, subjects will complete various questionnaires and tasks (e.g. Food Preference 
Questions, the Dutch Eating Behavior Scale, the Positive and Negative Affect Scale, hot flash 
diaries, reward seeking behavior, PSQI, ISI, PHQ-8). After waking, subjects may complete the post-
sleep questionnaire and the CCI IPM Exit Questionnaire, which are regularly administered by CCI 
inpatient unit staff. Subjects may also complete neurobehavioral testing administered automatically 
by the computers on the CCI inpatient unit. Throughout the inpatient portions of the study urine 
voids may be scheduled to be collected approximately every 3 hours when the subject is awake. 
Unscheduled urine voids during the day or overnight may also be collected. For each void, the urine 
volume will be measured, and aliquots may be saved for later hormonal, metabolite, or electrolyte 
analysis of analytes that may be affected by sleep fragmentation and/or changes in hormonal status. 
Urine samples will be collected and stored per standard techniques. Skin conductance may be 
measured using the Biolog skin conductance monitor.  During the 4 days prior to admission, subjects 
will receive several days of eucaloric meals from the CCI Nutrition Service, and ad lib breakfast 
will take place at BWH in order to measure food intake. Subjects will be instructed to standardize 
their sleep patterns during the 4 days at home, and up to a week beforehand, to extend the 
standardized routine.  
 Undisturbed procedures only: Bedtime and wake-up time will be fixed to an 8-hour standardized 
TST. 
 Fragmentation procedures only: During SBSF nights, subjects will be awakened using an auditory 
stimulus, adjusted to maintain the target number of awakenings and WASO observed in our most 
symptomatic subjects with leuprolide-induced HF (36 awakenings, 62 minutes WASO). TIB will 
be extended to maintain the 8-hour TST. Subjects will not be permitted to get out of bed or eat in 
the middle of the night. Auditory tones between 2 and 30-second duration and 60-110 dB will be 
used. Once awake, subjects will be required to stay awake for up to 2 minutes.  If subjects fall back 
asleep during the 1-2 minute awakening, the auditory stimulus will be played again to elicit an 
awakening. 
 
Pilot Study Procedures Overview 
 
To determine the most effective methods for fragmenting sleep in this protocol, we will first conduct 
a pilot study with up to 25 healthy volunteers. This pilot does not include intravenous lines or blood 
sampling procedures. These subjects will complete a much abbreviated version of the protocol 
detailed in the next section: 
A. Telephone screen: The Insomnia Severity Index (ISI) and the PHQ-2 will be asked over the 
phone. Those who meet initial eligibility criteria will be scheduled for an in-person screening 
visit. 
B. Screening visit: At the screening visit, written informed consent will be obtained and final 
eligibility criteria will be assessed, including a medical and psychiatric interview, sleep 

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interview, vitals, and a urine pregnancy test. All women who sign the consent form will be 
assigned a study identification number regardless of eligibility status. Eligible and consented 
participants will be scheduled for either a 1-night or 3-night stay in the Center for Clinical 
Investigation inpatient unit. 
C. Inpatient visit: Subjects will be admitted for a one-night or three-night stay within the BWH 
Clinical and Translational Science Center/Center for Clinical Investigation (CCI) inpatient 
unit. Subjects will be admitted in the afternoon and discharged mid-morning. During the stay, 
we will conduct the following abbreviated protocol: 
• 
On the day of admission, a urine pregnancy test will be performed, and vitals and 
morphometric measures will be gathered.  Subjects will remain on the 9B floor during the 
stay. Nursing staff will hook subjects up to a PSG monitor each night, and 24-hour skin 
conductance will be measured during the night using the Biolog skin conductance monitor. 
• 
Subject will be awakened during the night through auditory stimulus broadcast manually 
by sleep technicians throughout the night to achieve the target number of awakenings and 
minutes of Wake After Sleep Onset (WASO) (36 awakenings, 62 minutes WASO). Time 
In Bed (TIB) will be extended to maintain the 8-hour Total Sleep Time (TST). Auditory 
tones will be between 2- and 30-seconds duration, beginning at 60 dB and increased to a 
maximum of 1100db if an EEG awakening is not elicited. Once awake, subjects will be 
required to stay awake for up to 2 minutes. Subjects will not be permitted to get out of bed 
or eat in the middle of the night. 
• 
Subjects will be able to order meals and snacks off a provided menu. There will be no 
dietary restrictions. 
Nurses and technicians will be present 24 hours a day, so participants will have frequent social 
interactions in this controlled setting. Medical and psychologist/psychiatrist evaluation and 
treatment is available at all times. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

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Table of Procedures – Study overview 
 
 
 
Study Endpoints 
Metabolic endpoints are: 1) fasting morning levels of leptin (primary) adipokines, ghrelin (total, 
acetylated) (secondary), and 2) the circadian leptin profile (exploratory), calculated as area under 
curve (AUC) to detect whether the overall profile is lower, as seen in sleep restriction studies, and 
morning fasting AD levels. Behavioral endpoints are: 1) satiety (primary) and hunger (secondary) 
on a VAS scale, and 2) daily caloric intake during ad lib meals (exploratory). Analyses of each 
endpoint will be conducted in parallel as we seek to detect coherence among endpoints and focus 
on consistency across different outcomes in each hormonal and HF context. 
 
 
 
This study involves the administration of leuprolide to healthy individuals in order to better 
understand experimentally induced hot flashes. Subjects are healthy individuals who do not have 
any specific medical diagnosis. The DXA scan is not clinically indicated for women participating 
in the protocol.  Alternative imaging studies are therefore not indicated. 
For studies involving treatment or diagnosis, provide information about standard of care at 
Partners (e.g., BWH, MGH) and indicate how the study procedures differ from standard care. 
Provide information on available alternative treatments, procedures, or methods of diagnosis. 

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There will be a number of different procedures in place to minimize the risks to the participants. 
The PI’s current IND for use of leuprolide in healthy volunteers to study hot flashes (#73144) has 
been amended to include the current protocol. All study procedures will be approved by our IRB 
before being initiated. All adverse event reports will be reviewed by the PI. Serious and non-serious 
adverse events will also be reported to the Safety Monitoring Group (see DSMG form), FDA (for 
IND #73144) and our IRB in accordance with the respective reporting guidelines. Unanticipated 
problems involving risks to subjects or others including adverse events will be reported to the PHRC 
in accordance with PHRC unanticipated problems reporting guidelines. In the case of a serious 
adverse event, study procedures will be discontinued and appropriate diagnostic and therapeutic 
measures will be taken, as decided by the investigators. 
 
Thorough screening procedures will be implemented to exclude women in whom study treatments 
and procedures are contraindicated. Subjects participating in the study protocol will be monitored 
closely for the development of any adverse events. Criteria for withdrawing participants from the 
protocol in the event of an adverse event or serious health concern will be implemented. Women 
who develop a serious health concern or adverse event will be withdrawn from the protocol and 
referred for appropriate treatment. Potential risks to study subjects will be further minimized by the 
close monitoring and contact with study staff as required by the protocol design. 
 
All research records will be kept with maximum possible confidentiality. Results, including 
biological specimens, will be coded and the key to the code will be assigned by the PI and 
maintained in a locked file. No names will be used in presentation of any data. Source documents 
will be reviewed by study personnel. All HIPAA regulations will be strictly followed. 
 
The PI will meet with the project manager and research coordinator on a frequent basis to review 
and monitor the validity and integrity of all data in study binders and adherence to the IRB 
regulations. A checklist will also be included in the binders for each subject at each study visit. 
This checklist will be completed by the research coordinator and reviewed by the PI to confirm that 
all data are correctly obtained. In addition, all study personnel will review study data and discuss 
QA issues related to protocol adherence. Forms available on our IRB website will be used to keep 
track of adverse events and minor protocol violations. 
 
 
Participants will have a serum pregnancy test at the screening visit to ensure that they are not 
pregnant, and an additional urine pregnancy test will be conducted immediately prior to leuprolide 
administration. Subjects will be required to use a barrier method of contraception for the duration 
of the study and after study completion until menses resume. These procedures will serve to 
minimize the likelihood that study participants become pregnant during study procedures and during 
Describe how risks to subjects are minimized, for example, by using procedures which are 
consistent with sound research design and which do not unnecessarily expose subjects to risk or 
by using procedures already being performed on the subject for diagnostic or treatment purposes. 
Describe explicitly the methods for ensuring the safety of subjects. Provide objective criteria for 
removing a subject from the study, for example, objective criteria for worsening disease/lack of 
improvement and/or unacceptable adverse events. The inclusion of objective drop criteria is 
especially important in studies designed with placebo control groups. 

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the period of medication exposure. 
Participants will receive safety follow up calls regularly after study completion to ensure that 
menses resume and that, where applicable, that hot flashes resolve. If menstrual cycles do not return 
to normal or hot flashes do not resolve within several months of follow-up after the completion of 
all study procedures, participants be referred to an endocrinologist for further evaluation. We have 
used the same safety procedures in previous studies. 
 
Once in the laboratory, the subject will be constantly supervised by laboratory personnel. The 
medical physician for the BWH Clinical And Translational Science Center/Center for Clinical 
Investigation (CCI) inpatient unit will also be available on-call to handle any medical emergency 
that might arise. The subject’s vital signs (pulse rate, respiratory rate, systolic and diastolic blood 
pressure) will be checked at least once per day. An approximately 72” long intravenous line will be 
placed during each of the CCI inpatient unit admissions so that venous blood samples can be 
obtained using an IV line from outside the room without repeated venipuncture or sleep interruption 
related to the blood draw. If issues with the IV occur, then select samples, up to 2 per wake episode 
may be drawn via venipuncture. Prior to the insertion of the IV catheter, the skin around the insertion 
site will be washed with a Betadine solution followed by alcohol. An anti-bacterial ointment will 
be applied to the IV catheter insertion site prior to dressing the site. Hypoallergenic tape will be 
used on the IV dressing to minimize the development of contact dermatitis. The IV catheter insertion 
will be done while the subject is supine, and a staff member will remain in the suite with the subject 
for at least 10 minutes after the insertion to ensure that the subject does not feel light-headed or 
dizzy. Blood-sparing techniques will be used to minimize blood loss. The total amount of blood 
drawn will be up to 447.5 cc. Federal guidelines allow for up to 550 cc of venous blood per 2-month 
period in healthy non- pregnant adults. Anemic subjects will be excluded from participation and 
hemoglobin will be repeated at the completion of the study to confirm the absence of anemia. Oral 
ferrous sulfate supplementation and clinical follow up will be provided if anemia is present. 
 
The PI has established additional monitoring procedures for specific adverse events related to use 
of leuprolide in healthy volunteers which will be implemented in the protocol. While there is a low 
likelihood (2% in our experience) that a subject will develop significant depressive symptoms, 
mood will be assessed repeatedly throughout the study as part of standard safety monitoring 
procedures. Women developing severe depression or who have suicidal or homicidal ideation will 
be withdrawn from the study and referred for appropriate care. If suicidal or homicidal ideation is 
of concern, subjects will be evaluated immediately. Dr. Joffe is an experienced psychiatrist with an 
expertise in women’s mental health who will be available to evaluate and treat women with 
significant depression. 
 
Off-Study Criteria: 
1. Emergence of significant psychiatric symptoms, suicidal or homicidal ideation 
2. Initiation of hormonal or centrally acting medications 
3. Development of any significant medical problem 
4. Pregnancy 
5. Significant deviations from study protocol, including enrollment in another research protocol that 
may influence the results of the current protocol. 
 
 

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FORESEEABLE RISKS AND DISCOMFORTS 
 
Potential risks associated with study medications: 
Leuprolide in healthy volunteers: Leuprolide is a widely used gonadotropin releasing hormone 
(GnRH) agonist that is manufactured by Abbott. Leuprolide is indicated for treatment of 
endometriosis, uterine fibroids, precocious puberty, and prostate cancer, and has been used off- 
label for in-vitro fertilization and premenstrual syndrome. In the current protocol, leuprolide will be 
administered at a dose routinely used for treatment of endometriosis and uterine fibroids in 
premenopausal women. Leuprolide can induce amenorrhea for 2 to 3 months after a depot injection 
is given. However, unpredictable menstrual bleeding and spotting can occur during this time period. 
Menstrual-cycle patterns will be monitored closely during the study. Through its effects on female 
hormones, leuprolide temporarily reduces fertility for up to 3 months after a depot injection is given. 
Leuprolide may cause growth abnormalities and death in an unborn fetus. 
Although long-term administration of any GnRH agonist (i.e., >6 months) is associated with bone 
loss, bone effects are not expected in this protocol because only a single dose of GnRH agonist 
therapy will be administered. Other side effects are related to suppression of ovarian steroid 
secretion, including: 
Adverse reactions to short-term use: 
 Common: Hot flashes, headache, vaginal bleeding, vaginal dryness, vaginitis, alopecia, body 
and joint pain, emotional lability, depression symptoms, memory problems, nausea, vomiting, 
peripheral edema 
 Rare (<3%): Myalgia, hirsutism, acne, weight gain or weight loss, depression, decreased libido, 
dyspareunia, paresthesia, skin reactions, dizziness, breast tenderness and pain 
Leuprolide is associated with emotional lability in 22–31% of women receiving leuprolide as part 
of large clinical trials for treatment of endometriosis and fibroids. These studies do not provide 
information about whether the mood symptoms that emerged were persistent or at a clinically 
significant level suggesting a depressive disorder. However, our experience, and that of other 
reproductive psychiatry investigators using close monitoring of mood in healthy volunteers given 
leuprolide, indicates that clinically significant depression is rare (2–3%). Additional information 
from the manufacturers of leuprolide also indicates that serious depression develops only rarely on 
GnRH agonists.Potential risks associated with DXA scan: A dual-energy x-ray absorptiometry 
(DXA) scan will be performed to quantify total body fat mass. The total body DXA scan is non-
invasive and takes less than 10 minutes to complete with minimal/no discomfort while lying down 
on a scan table under the supervision of the Bone Density staff. Participants will be exposed to a 
small amount of radiation from this scan. The amount of radiation exposure with DXA is even less 
than that of an abdominal CT scan. The exposure from the DXA scans is equal to less than 1% of 
the dose permissible during occupational exposure. The risk from radiation exposure of this level is 
too small to be measured directly and is considered to be negligible in comparison with other 
everyday risks. Pregnancy tests will be used to ensure that women undergoing the DXA scans are 
not pregnant. 
Patients will be screened for contraindications to DXA as part of the eligibility/exclusion 
requirements so that they do not participate in this protocol. 
Provide a brief description of any foreseeable risks and discomforts to subjects.  Include those 
related to drugs/devices/procedures being studied and/or administered/performed solely for 
research purposes.  In addition, include psychosocial risks, and risks related to privacy and 
confidentiality.  When applicable, describe risks to a developing fetus or nursing infant. 

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It is possible that the DXA scan may coincidentally detect an abnormality that requires further 
evaluation. The participant and her physician (with the subject’s permission) will be informed of 
the scan results so follow-up assessments and treatment can be undertaken if necessary. 
 
Potential risks associated with Biolog Ambulatory Hot Flash Monitor: The Biolog ambulatory 
hot flash monitor (UFI, Morro Bay, CA) will be used to measure hot flashes objectively. There are 
no significant risks associated with skin-conductance methodologies. The input assembly wiring is 
fixed so that the subject is inherently isolated, making the risk of current leakage negligible. Skin 
conductance is measured by 2 adhesive gel electrodes that are attached to the sternum. There is a 
minor risk of skin irritation from the adhesive gel. The recorder and electrodes cannot get wet; 
therefore subjects will be instructed to not bathe while wearing the monitor. 
 
Potential risks associated with inpatient sleep study procedures: Subjects will be admitted to the 
BWH Clinical And Translational Science Center/Center for Clinical Investigation (CCI) inpatient 
unit for 5 nights at a time. Admissions of this length may be psychologically stressful and 
inconvenient or disruptive to daily routines. Headache, nausea, dizziness, light headedness, 
excessive sleepiness, muscle soreness, and weakness may occur. During some of the admissions, 
subjects will experience experimentally induced sleep fragmentation which will reduce their quality 
of sleep, as a minor discomfort. Short term sleep fragmentation can cause drowsiness and increase 
the risk of accidents. While subjects will be monitored and accidents are unlikely in the controlled 
environment of the laboratory, we have less control when subjects leave the laboratory. Thus, we 
will ensure that the subject fully appreciates this risk and has safe transportation home from the unit 
after each of the sleep fragmentation sleep blocks. The subjects may become sleepy during some 
segments of the study but will be asked to remain awake during the entirety of their scheduled wake 
times. Should they feel unable to remain awake, they are free to withdraw consent to participate in 
this experiment and then go to sleep. After the sleep fragmentation procedures, subjects may have 
some difficulty sleeping and waking at their usual times for several days. Volunteer subjects are 
studied in the CCI inpatient unit within the main BWH campus. In the unlikely event that emergency 
treatment is required during the subject’s stay in the research facility, treatment will be provided. 
 
Polysomnography: PSG studies will be conducted according to standard clinical procedures, which 
have minimal risks. A minor rash may develop from the tape or adhesives used to attach electrodes, 
but this is expected to resolve within a few days. In addition, the monitoring equipment may make 
sleep somewhat more difficult than usual and the individual may therefore experience minimally 
reduced quality of sleep. 
 
Ambulatory Polysomnography: The ambulatory polysomnography equipment is commonly used 
in clinical sleep testing and has minimal risk. A minor rash may develop from tape or adhesives 
used to attach the electrodes, but is expected to resolve within a few days. In addition, the monitoring 
equipment may make sleep somewhat more difficult than usual and the individual may therefore 
experience minimally reduced quality of sleep.  
 
Potential risks associated with venous blood sampling: The risks associated with venipuncture and 
intravenous lines are minimal and include hematoma, pain, infection, and fainting spells. All blood 
draws and IV placements will be performed by trained personnel that use standard sterile 
techniques. Infection is unlikely since stringent aseptic techniques are followed during all 

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procedures. An approximately 72” long intravenous line will be placed for a 24-hour period during 
each of the BWH Clinical And Translational Science Center/Center for Clinical Investigation (CCI) 
inpatient unit so that venous blood samples can be obtained up to every hour using an IV line from 
outside the room without repeated venipuncture or sleep interruption related to the blood draw. 
Blood-sparing techniques will be used to minimize blood loss. If issues with the IV occur, then 
select samples, up to 2 per wake episode may be drawn via venipuncture. There may be some side 
effects from the use of heparin; such as bleeding and allergy. 
There may be some discomfort or bruising on initial insertion of the catheter into a vein, but wearing 
the catheter should not be painful. Occasionally, mild discomfort may occur from the tube in the 
vein. If this happens, it can be repositioned or removed, asking the subject’s permission before any 
subsequent reinsertion. Occasionally, there is a bruise at the site of the IV insertion, which may last 
a few weeks; and, rarely, a small scar may remain permanently at the venipuncture site. There may 
be a minor skin rash or reaction to the sterile tape (contact dermatitis) used to hold the catheter in 
position. 
A total of up to 447.5 cc of venous blood will be drawn throughout the study. Federal guidelines 
allow for up to 550 cc of venous blood per 2-month period in healthy non-pregnant adults weighing 
>110 pounds. There is a risk of becoming anemic from frequent blood sampling. 
 
Psychological risks: Answering questions on the research instruments and questionnaires used to 
evaluate psychological symptoms can be upsetting to some women. Subjects will be told that they 
can skip specific questions that make them uncomfortable. Any woman who develops severe 
depression during the study will be evaluated and referred for treatment, including emergency 
evaluation if appropriate. 
 
Radiation exposure: Each set of DEXA scans result in an effective radiation dose of approximately 
0.5 mrem so that both sets results in an effective dose of approximately 1 mrem. The total effective 
dose is therefore approximately 1 mrem.   
 
Risks of MealLoggerTM: The research team will create credentials for patients using this app, so 
that MealLogger will not be able to associate the data with a real person. It is not necessary for the 
study staff to collect identifiable information through the app, so participants will be instructed that 
the only required information is their meals. MealLogger is easily installed on both Android and 
iPhone systems; the research team does not anticipate usability issues given that MealLogger is used 
both in research studies and in the general populations, but participants will be instructed to contact 
research coordinators with any questions about the technology. MealLogger has already been 
approved in other Brigham and Women’s Hospital research studies (e.g., 2012P-001631). 
 
Risks of Study Food: The study food provided by the BWH CCI Dietary Core is specific to each 
participant’s preferences and restrictions. The Dietary staff are informed of participants’ allergies, 
as well as preferences (e.g., vegetarian or vegan). There is a minor risk of gastrointestinal symptoms 
(e.g., stomach discomfort/ache, nausea, or abdominal bloating) as participants’ diets at home may 
not include the same types of food consumed during the study diet.  
 
 
 
 
 

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EXPECTED BENEFITS 
 
Benefits to subjects 
As healthy volunteers, participants will not receive any direct benefit from this study protocol. 
However, the protocol is expected to provide a meaningful benefit to peri- and postmenopausal 
women through knowledge gained about the metabolic consequences of sleep disruption, hot 
flashes, and loss of circulating estradiol. For those participating in the study, the sleep studies, 
radiographic assessments, morphometric measures, metabolic and reproductive hormone levels 
may be of interest. If a participant requests routine health information obtained during the study, 
she will be provided with a copy after completing study procedures. If an abnormality is detected 
during the study, this information will be shared with the participant and her physician (with her 
permission). Our experience to date indicates that some women find the knowledge obtained 
through the experience of developing transient menopausal symptoms informative. Because there 
is no direct benefit of study procedures to these healthy volunteers, they will be compensated for 
their time, effort and willingness to undergo study procedures, including exposure to time-limited 
leuprolide therapy and time spent in the BWH Clinical And Translational Science Center/Center 
for Clinical Investigation (CCI) inpatient unit. However, financial compensation to subjects will not 
be presented as a benefit of participation in this research. 
 
Importance of the knowledge to be gained 
Obesity is highly prevalent in midlife and older women, with rates increasing markedly after age 40 
and abdominal adiposity increasing in half of women during and after the menopause transition 
independent of age, but the factors that cause these changes are not well understood. Reduced total 
sleep time has been shown to adversely impact biomarkers of obesity, but the effect of menopause-
related sleep fragmentation, which occurs in >40% of midlife women and is characterized by sleep 
interruption secondary to hot flashes, on metabolism and eating behaviors is not known. The broad 
goal of this proposal is to isolate the impact of menopause-related sleep disruption, as well as 
menopause-related hot flashes and estrogen withdrawal, on behavioral and metabolic biomarkers of 
body fat gain, results of which will inform strategies to prevent gains in body fat, thereby improving 
cardiovascular and metabolic health outcomes in women. 
 
EQUITABLE SELECTION OF SUBJECTS 
 
 
The study population is restricted to non-pregnant premenopausal women, the population for whom 
Describe both the expected benefits to individual subjects participating in the research and the 
importance of the knowledge that may reasonably be expected to result from the study. Provide 
a brief, realistic summary of potential benefits to subjects, for example, “It is hoped that the 
treatment will result in a partial reduction in tumor size in at least 25% of the enrolled subjects.” 
Indicate how the results of the study will benefit future patients with the disease/condition being 
studied and/or society, e.g., through increased knowledge of human physiology or behavior, 
improved safety, or technological advances. 
The risks and benefits of the research must be fairly distributed among the populations that stand 
to benefit from it.  No group of persons, for example, men, women, pregnant women, children, 
and minorities, should be categorically excluded from the research without a good scientific or 
ethical reason to do so.  Please provide the basis for concluding that the study population is 
representative of the population that stands to potentially benefit from this research. 

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the information obtained in this study is relevant. Pregnant women are excluded because of risks 
with study medication to the unborn fetus. Children under 18 are excluded because the study is not 
relevant to pre-pubertal children and the hypothalamic-pituitary-ovarian axis is in flux in many post-
pubertal teenage girls, as reflected by frequent menstrual-cycle abnormalities in this population. 
The age requirements are that women must be within 18-45 years of age in order to study 
experimentally induced hot flashes in women who are close to natural menopause. Men are 
excluded because the hormonal changes (e.g., estrogen) that occur with GnRH agonists differ 
between women and men and these outcomes are important components of this study. Minority 
populations are included. 
Individuals who do not speak English will be excluded from participation in this study because 
many of the questionnaires administered during this study are not validated in other languages. 
Participants will be recruited from outpatient primary care departments at BWH and affiliated 
community clinics as well as through the larger Boston community, using methods we have 
employed to successfully recruit healthy volunteers to similar studies. Advertising methods include 
using the Research Study Volunteer Program (RSVP for Health), an online system through which 
registered individuals receive information about research studies, recruitment postcards, and paid 
newspaper and radio advertisements to publicize the study. Study staff may also use other internet 
forums (e.g., Craigslist, Facebook) to recruit. Women inquiring about the study will be screened 
preliminarily over the phone by the research coordinator using an IRB-approved telephone script, 
and if appropriate for the study, will be invited for a 2-part screening visit: Visit 1a will be remote 
and will include electronic signatures for written consent via REDCap e-consent template obtained 
by a study clinician, and Visit 1b will be in-person for a brief physical exam and blood draw. 
Study participants will be compensated for the time, effort, and study procedure risks involved in 
this intensive protocol. Participants will receive up to $8,055 by check, upon completing the 
study. We anticipate that some women will not be eligible after completing screening procedures 
or will decline to continue to participate after Sleep Block 1.  
When people who do not speak English are excluded from participation in the research, provide 
the scientific rationale for doing so.  Individuals who do not speak English should not be denied 
participation in research simply because it is inconvenient to translate the consent form in 
different languages and to have an interpreter present. 
Explain in detail the specific methodology that will be used to recruit subjects.  Specifically 
address how, when, where and by whom subjects will be identified and approached about 
participation.  Include any specific recruitment methods used to enhance recruitment of women 
and minorities. 
RECRUITMENT PROCEDURES
Provide details of remuneration, when applicable.  Even when subjects may derive medical 
benefit from participation, it is often the case that extra hospital visits, meals at the hospital, 
parking fees or other inconveniences will result in additional out-of-pocket expenses related to 
study participation.  Investigators may wish to consider providing reimbursement for such 
expenses when funding is available 

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Subjects will be paid at end of the study and will accumulate compensation according to the 
following schedule. 
Screening visits: up to $100 ($25 for completing Visit 1a and $75 for completing Visit 1b) 
DXA Scan: $55 
Lupron Administration Visit: $250 
Inpatient data collection: $275 per night spent in lab 
Outpatient data collection: up to $700 per Sleep Block ($50/day; up to 7 days prior to and 7 days 
after each Sleep Block) 
Bonus for completion of all study visits and procedures: $3500 
If subjects do not complete the study, they will be paid for the portion of the study completed but 
will not receive any study bonus.  
Subjects will not have to pay for any meals during their inpatient stays. Reimbursement for 
transportation home after each of the 2 inpatient visits will be reimbursed up to $25 each with 
receipts (total up to $50). Valet parking vouchers will be provided for outpatient visits. 
Subjects in the pilot study will be compensated $50 for the screening visit and $250 for each night 
they stay in the unit (maximum compensation = $800). 
CONSENT PROCEDURES 
Subjects will not undergo any study procedures until it is clear that they understand the risks and 
benefits and consent to participate. At the remote screening visit (V1a), a research physician will 
review the consent form with prospective subjects. Women enrolling in the study must be capable 
of understanding the nature of the study as well as the discomforts and potential benefits. Any 
questions or areas of concern regarding the protocol or alternative treatment options will be 
addressed by one of the physician investigators. If a woman feels comfortable with the study 
procedures, she will sign the consent form along with the physician investigator using the electronic 
REDCap e-consent template. The consent form will include information about the risks and benefits 
of study participation and contact information for the study physicians and research staff. The study 
protocol, consent forms, telephone screens, and all other study procedures will be approved by the 
IRB before the study is initiated. 
Enrolled subjects will be given a copy of the completed consent form before any study procedures 
are completed, and a copy of the signed consent form will also be placed in the subject’s research 
file. If subjects have reservations about study participation at the first visit, they have the option to 
take the consent form home to review the decision to participate in the study again with research 
physicians before signing the consent form and initiating study procedures. 
Additional considerations: Because of the nature of drug administration to healthy volunteers, the 
Explain in detail how, when, where, and by whom consent is obtained, and the timing of consent 
(i.e., how long subjects will be given to consider participation). For most studies involving more 
than minimal risk and all studies involving investigational drugs/devices, a licensed physician 
investigator must obtain informed consent.  When subjects are to be enrolled from among the 
investigators’ own patients, describe how the potential for coercion will be avoided. 

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informed consent process will include a discussion about the use of leuprolide for FDA- approved 
indications in premenopausal women (e.g., endometriosis, fibroids). It will be made clear that study 
participants will have no direct health benefit from the treatment because they have no medical 
reason to receive the treatment. Participants will be informed that the motivation to administer a 
GnRH agonist to healthy volunteers is to understand metabolic consequences of sleep fragmentation 
associated with hot flashes induced by leuprolide, as well as that of hot flashes and estradiol 
withdrawal. It will be explained that a research participant retains the right to withdraw from a study 
at any point, but that the effects of the GnRH agonist on gonadal hormones cannot be reversed if 
she chooses to withdraw after receiving leuprolide because leuprolide is being administered in a 
depot form. Participants will also be informed that the study procedures are being conducted with 
the approval and monitoring of the IRB and FDA. They will also be told that if any abnormalities 
are detected as part of study procedures (e.g., on the DXA scan), results of these tests will be relayed 
to the subject and her physician (with her permission) so that follow up assessments and treatment 
can be undertaken if necessary. 
 
DATA AND SAFETY MONITORING 
 
We will continue to use the Safety Monitoring Group (SMG) that we set up for prior studies using 
leuprolide in healthy volunteers, consistent with regulatory guidelines. This Group will continue to 
serve as the SMG for the proposed project as well, meeting approximately every 6 months to review 
the progress of the study. The Group comprises two research physicians at Massachusetts General 
Hospital, Harvard Medical School, who are familiar with clinical research and the study medications 
and their side effects, but they are not collaborators or close colleagues of the PI or co-investigators. 
The two physicians are a neuro-endocrinologist (Karen Miller, MD) and a psychiatrist/ neurologist 
(John Winkelman, MD, PhD). Drs. Miller and Winkelman will review all adverse events and 
unanticipated problems that arise during the study. Adverse events, including serious adverse 
events, and unanticipated problems will be managed and reported to our local Institutional Review 
Board consistent with their guidelines. We have also amended Dr. Joffe’s IND (#73144) to link it 
with this study and will continue to submit annual reports, including IRB updates and SMG minutes, 
to the FDA as part of the safety monitoring of this IND. 
 
Describe the plan to be followed by the Principal Investigator/study staff for review of adverse 
events experienced by subjects under his/her care, and when applicable, for review of sponsor safety 
reports and DSMB reports.  Describe the plan for reporting adverse events to the sponsor and the 
Partners’ IRB and, when applicable, for submitting sponsor safety reports and DSMB reports to the 
Describe the plan for monitoring the data to ensure the safety of subjects.  The plan should 
include a brief description of (1) the safety and/or efficacy data that will be reviewed; (2) the 
planned frequency of review; and (3) who will be responsible for this review and for determining 
whether the research should be altered or stopped.  Include a brief description of any stopping 
rules for the study, when appropriate.  Depending upon the risk, size and complexity of the 
study, the investigator, an expert group, an independent Data and Safety Monitoring Board 
(DSMB) or others might be assigned primary responsibility for this monitoring activity. 
 
NOTE: Regardless of data and safety monitoring plans by the sponsor or others, the principal 
investigator is ultimately responsible for protecting the rights, safety, and welfare of subjects 
under his/her care. 

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Partners’ IRBs.  When the investigator is also the sponsor of the IND/IDE, include the plan for 
reporting of adverse events to the FDA and, when applicable, to investigators at other sites. 
 
NOTE: In addition to the adverse event reporting requirements of the sponsor, the principal 
investigator must follow the Partners Human Research Committee guidelines for Adverse Event 
Reporting 
Subjects will be closely monitored for adverse events at every visit. Research staff will record any 
serious or non-serious adverse events using standard spontaneous IRB-approved reporting 
procedures. Participants will be informed that they can contact the investigator between visits if 
they have concerns about potential side effects. Routine adverse events and serious adverse events 
will be reported to the FDA and Human Research Committee in accordance with the FDA and 
Human Research Committee reporting guidelines, following the timeframes specified by the 
Partners Investigator’s Guidelines. In addition, the DSMG will review all adverse events and 
investigate those considered serious and unexpected consistent with standard ISM procedures. In 
case of a serious adverse event, study procedures will be discontinued and appropriate diagnostic 
and therapeutic measures will be taken, as decided by the investigator. All adverse event reports 
will be reviewed by the principal investigator. 
 
MONITORING AND QUALITY ASSURANCE 
 
The PI will meet with the study coordinator on a weekly basis to review and monitor the validity 
and integrity of all data in study binders and adherence to the IRB-approved protocol. A checklist 
will also be included in the binders for each subject at each study visit.  This checklist will be 
completed by the research coordinator and reviewed by the PI to confirm that all data are collected.   
In addition, all study personnel will meet on a monthly basis to review study data and discuss QA 
issues related to protocol adherence.  Forms available on the IRB website will be used to keep track 
of adverse events and minor protocol violations. 
 
 
 
 
 
 
Describe the plan to be followed by the principal investigator/study staff to monitor and assure 
the validity and integrity of the data and adherence to the IRB-approved protocol. Specify who 
will be responsible for monitoring, and the planned frequency of monitoring.  For example, 
specify who will review the accuracy and completeness of case report form entries, source 
documents, and informed consent. 
 
NOTE: Regardless of monitoring plans by the sponsor or others, the principal investigator is 
ultimately responsible for ensuring that the study is conducted at his/her investigative site in 
accordance with the IRB-approved protocol, and applicable regulations and requirements of the 
IRB. 

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PRIVACY AND CONFIDENTIALITY 
 
Confidentiality will be maintained by storing data with a subject identification number only. Only 
the PI, co-investigators, and study staff will have access to the database and the study binders. Data 
will be entered into a secure, HIPAA-compliant Research Electronic Data Capture (REDCap) 
database hosted by Partners HealthCare and analyzed using STATA 14.0 (College Station, Texas). 
All exported data will be de-identified using only the subject’s identification number. DXA data 
will be reviewed by experts in bone densitometry and a radiologist, respectively. Analysis of all 
study data will be conducted by co-investigators/project leaders (MD/PhD level) under the purview 
of Dr. Joffe in consultation with Sybil Crawford, PhD, the statistical consultant on this project. 
 
 
SENDING 
SPECIMENS/DATA 
TO 
RESEARCH 
COLLABORATORS 
OUTSIDE 
PARTNERS 
 
De-identified data will be sent to our statistical consultant working with us at University of 
Massachusetts Medical School, Dr. Sybil Crawford.  Deidentified lab specimens will be sent to Dr. 
Cecilia Hillard at Medical College of Wisconsin and the NSF International laboratory for assays.  
NSF International is not considered a collaborator; rather, they are fee-for-service. All study 
materials will be labeled with a de-identified study ID only. No information that could be used to 
link the data to an individual subject will be sent to Dr. Crawford, Dr. Hillard, or NSF. 
 
 
 
No data or specimens will be stored at collaborating sites outside partners. Participants will be asked 
for permission to store serum samples and health information for future research related to 
menopause and sleep disturbance at Brigham and Women’s Hospital.  Participants may withdraw 
their consent to store samples at any time. 
 
 
Describe methods used to protect the privacy of subjects and maintain confidentiality of data 
collected.  This typically includes such practices as substituting codes for names and/or medical 
record numbers; removing face sheets or other identifiers from completed 
surveys/questionnaires; proper disposal of printed computer data; limited access to study data; 
use of password-protected computer databases; training for research staff on the importance of 
confidentiality of data, and storing research records in a secure location. 
 
NOTE: Additional measures, such as obtaining a Certificate of Confidentiality, should be 
considered and are strongly encouraged when the research involves the collection of sensitive 
data, such as sexual, criminal or illegal behaviors. 
Specimens or data collected by Partners investigators will be sent to research collaborators 
outside Partners, indicate to whom specimens/data will be sent, what information will be sent, 
and whether the specimens/data will contain identifiers that could be used by the outside 
collaborators to link the specimens/data to individual subjects. 
Specifically address whether specimens/data will be stored at collaborating sites outside 
Partners for future use not described in the protocol.  Include whether subjects can withdraw 
their specimens/data, and how they would do so. When appropriate, submit documentation 
of IRB approval from the recipient institution. 

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RECEIVING SPECIMENS/DATA FROM RESEARCH COLLABORATORS OUTSIDE 
PARTNERS 
 
We will not be receiving specimens or data collected by research collaborators outside Partners. 
When specimens or data collected by research collaborators outside Partners will be sent to 
Partners investigators, indicate from where the specimens/data will be obtained and whether the 
specimens/data will contain identifiers that could be used by Partners investigators to link the 
specimens/data to individual subjects.  When appropriate, submit documentation of IRB 
approval and a copy of the IRB-approved consent form from the institution where the 
specimens/data were collected.