Lifestyle Enriching Activities for Research in Neuroscience Intervention Trial: LEARNit Study
Phase: NA
Conditions: Aging, Cognitive Ability, General
Sponsor: University of Southern California

LEARNit Protocol 
Version 3.0 
April 26, 2024 
Lifestyle Enriching Activities for Research in Neuroscience – intervention trial  
Study Protocol 
 
NCT#:  
NCT0272690

LEARNit Protocol 
Version 3.0 
April 26, 2024 
 
Confidentiality Statement: 
  
 
 
 
The information in this document is privileged or confidential and may not be disclosed unless 
such disclosure is required by applicable laws and regulations. It should not be copied by or 
distributed to persons not involved in the study. Persons to whom the information is disclosed 
must be informed that the information is privileged or confidential and may not be further 
disclosed by them. These restrictions on disclosure will apply equally to all future information 
supplied to you, which is indicated as privileged or confidential. Any unauthorized review, use, 
disclosure, copying or distribution is strictly prohibited.  
 
  
 
 
 
  
 
 
  
 
 

 
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FULL PROTOCOL TITLE 
 
Lifestyle Enriching Activities for Research in Neuroscience intervention trial (LEARNit), 
a randomized, single blind, 60 participant clinical trial examining the effect of aerobic 
exercise vs. health education on brain function and cognition in older adults with early 
mild cognitive impairment. 
 
Principal Investigator: 
Judy Pa, Ph.D., University of Southern California 
 
Sponsor: 
The National Institute on Aging 
1R01AG046928 - 01A1 
 
Investigational PET ligand F-AV-1451 Provided by:  
Avid Pharmaceuticals  
 
IND holder:  
Dr. Peter Conti 
IND # 129822 
 
NCT#:  
NCT02726906
 
 
 

 
 
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TABLE OF CONTENTS 
FULL PROTOCOL TITLE ......................................................................................................... 1 
PRÉCIS ........................................................................................................................................... 5 
Study Title ................................................................................................................................... 5 
Objectives .................................................................................................................................... 5 
Interventions and Duration .......................................................................................................... 6 
Sample Size and Population ........................................................................................................ 6 
STUDY TEAM ROSTER ............................................................................................................. 7 
○ 
PI:    Judy Pa, PhD ............................................................................................................... 7 
○ 
Co-Investigators: Peter Conti, MD ...................................................................................... 7 
1 
Study objectives ..................................................................................................................... 9 
1.1 
Primary Objective ............................................................................................................ 9 
1.2 
Secondary Objectives ...................................................................................................... 9 
2 
BACKGROUND AND RATIONALE ................................................................................. 9 
2.1 
Background on Condition, Disease, or Other Primary Study Focus ............................... 9 
2.2 
Study Rationale .............................................................................................................. 10 
3 
STUDY DESIGN ................................................................................................................. 10 
4 
SELECTION AND ENROLLMENT OF PARTICIPANTS ........................................... 11 
4.1 
Inclusion Criteria ........................................................................................................... 11 
4.2 
Exclusion Criteria .......................................................................................................... 11 
4.3 
Study Enrollment Procedures ........................................................................................ 12 
5 
STUDY INTERVENTIONS ............................................................................................... 13 
5.1 
Interventions, Administration, and Duration ................................................................. 13 
5.2 
Handling of Study Interventions .................................................................................... 13 
5.3 
Concomitant Interventions ............................................................................................ 16 
5.3.1 
Allowed Interventions ........................................................................................... 16 
5.3.2 
Required Interventions .......................................................................................... 16 
5.3.3 
Prohibited Interventions ........................................................................................ 16 
5.4 
Adherence Assessment .................................................................................................. 16 

 
 
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5.5 
Schedule of Evaluations .................................................................................................. 18 
6 
Description of Evaluations ................................................................................................ 19 
6.1 Screening Evaluation ....................................................................................................... 19 
6.1.1 
Enrollment, Baseline, and/or Randomization ........................................................ 19 
6.1.2 
Follow-up Visits / Completion/Final Evaluation ................................................... 20 
7 
SAFETY ASSESSMENTS .................................................................................................. 21 
7.1 
Adverse Events and Serious Adverse Events ................................................................ 21 
7.2 
Reporting Procedures .................................................................................................... 22 
7.3 
Follow-up for Adverse Events ....................................................................................... 22 
7.4 
Safety Monitoring .......................................................................................................... 22 
8 
INTERVENTION DISCONTINUATION ........................................................................ 22 
9 
STATISTICAL CONSIDERATIONS ............................................................................... 23 
9.1 
General Design Issues ................................................................................................... 23 
9.2 
Sample Size and Randomization ................................................................................... 23 
9.2.1 
Treatment Assignment Procedures ........................................................................ 24 
9.3 
Interim analyses and Stopping Rules ............................................................................. 24 
9.4 
Outcomes ....................................................................................................................... 24 
9.4.1 
Primary outcome .................................................................................................... 24 
9.4.2 
Secondary outcomes .............................................................................................. 25 
9.5 
Data Analyses ................................................................................................................ 25 
10 
DATA COLLECTION AND QUALITY ASSURANCE ............................................. 26 
10.1 
Data Collection Forms ................................................................................................... 26 
10.2 
Data Management .......................................................................................................... 26 
10.3 
Quality Assurance .......................................................................................................... 26 
10.3.1 
Training ................................................................................................................. 26 
10.3.2 
Quality Control Committee ................................................................................... 26 
10.3.3 
Metrics ................................................................................................................... 27 
10.3.4 
Protocol Deviations ............................................................................................... 27 
10.3.5 
Monitoring ............................................................................................................. 27 
11 
PARTICIPANT RIGHTS AND CONFIDENTIALITY .............................................. 27 
11.1 
Institutional Review Board (IRB) Review .................................................................... 27 
11.2 
Informed Consent Forms ............................................................................................... 27 

 
 
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11.3 
Participant Confidentiality ............................................................................................. 27 
11.4 
Study Discontinuation ................................................................................................... 28 
12 
ETHICAL CONSIDERATIONS ................................................................................... 28 
13 
PUBLICATION OF RESEARCH FINDINGS ............................................................. 29 
REFERENCES ............................................................................................................................ 30 
 
 
SUPPLEMENT / APPENDIX 
 
Appendix I - Informed Consent Form 
 
Appendix II - Covid - 19 Required Safety Protocols 
 
 

 
 
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PRÉCIS 
Alzheimer’s disease (AD) is the most common cause of cognitive impairment in older adults.1 
The need for effective treatments has become imperative, as 36 million people worldwide are 
affected.2,3 The results from several large trials have been sobering with a lack of significant 
clinical outcomes, prompting a paradigm shift to initiate treatment earlier in the disease process 
and explore lifestyle interventions.4-7 Exercise has emerged as an exciting, non-pharmacological 
approach to help remediate cognitive loss or delay onset of dementia.8-13 However, to fully 
capitalize on the investments in exercise and to discover the full treatment potential of exercise, 
the brain mechanisms supporting exercise for older adults with MCI must be understood.  
 
Using an aerobic walking intervention program, we will conduct an investigation of brain 
mechanisms supporting exercise, from localized function and structure of the hippocampus, to 
lower-level growth factors. This project will recruit 60 sedentary older adults between 55-80 
years of age with early MCI. Based on the Surgeon General’s recommendation for moderate-
intensity aerobic in older adults,14-15 participants will be randomly assigned to: aerobic walking 
for 30 mins, 5 days/week for 6 months OR health education non-active control for 6 months, 
with 30 participants in each arm. Primary outcome measurements include regional brain activity 
in the hippocampus and behavioral performance on the Mnemonic Similarity Task (MST).16-17 
Additionally, we will examine secondary outcomes of physical function, aerobic fitness, amyloid 
burden, and brain derived neurotrophic factor (BDNF) associated with exercise.18-21 For 
exploratory purposes, all participants will be scanned with amyloid and tau PET tracers at 
baseline and follow-up to examine the contribution of amyloid and tau aggregation on brain 
function and to serve as an AD risk biomarker.  
  
Study Title  
Lifestyle Enriching Activities for Research in Neuroscience intervention trial (LEARNit)  
Objectives  
The purpose of this study is to examine the effects of modifiable lifestyle factors 
including exercise and healthy living education on brain health. The investigators will compare 2 
types of interventions, moderate aerobic walking vs. healthy living education reading, over 6 
months to evaluate changes in brain function, cognition, and physical function in older adults 
with cognitive concerns.   
 
 
 
 
 
Our primary outcomes are neuroimaging measures of brain function after a 6--month exercise 
intervention, including regional BOLD activity and performance on a memory task, with 
secondary outcomes of brain derived neurotrophic factor, amyloid PET, physical function and 
aerobic fitness. We will link biological changes to meaningful clinical outcomes by examining 

 
 
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the relationship between enhanced brain function, improved memory performance and physical 
function after the intervention. 
 
 
 
Primary outcomes. Based on the goals and innovation of this proposal, our primary outcomes are 
neuroimaging measure of brain function: fMRI BOLD signal in the hippocampus. Functional 
MRI has proven to be an effective non-invasive, in-vivo tool for measuring brain function. 22-24 
Growing evidence supports that fMRI BOLD can detect neuronal changes that occur very early 
in the disease process. The hippocampus has been shown to be a selectively vulnerable region in 
AD, and several studies, including our own, identified a dysfunctional brain signal in the 
hippocampus in MCI patients.25-27  
 
Secondary outcomes. The secondary outcomes consist of additional biological and clinical 
measures. The clinical outcomes were selected based on previous literature examining the effects 
of exercise and for enabling comparisons to the large ADCS exercise trial. The cognitive and 
functional outcome measures will be measured pre- and post-intervention (at baseline and 6 
month follow-up).  
 
Design and Outcomes   
 
This trial is a single-blind, randomized controlled trial to test the efficacy of aerobic exercise on 
brain signal and memory function in individuals 55-80 years of age.  
Interventions and Duration  
There will be two arms:  
 
Moderate aerobic walking OR Health education reading 
 
Participants will engage in the arm specific intervention activities for the duration of the 6-month 
intervention. There will be two study visits, one at baseline and one at the end of the 6-month 
intervention.  
Sample Size and Population  
LEARNit will recruit 60 sedentary older adults ages 55-80 displaying early signs of cognitive 
impairment, as identified by the cognitive assessments administered at the screening visit via 1 
SD or lower on one cognitive test from a neuropsychological battery. Subjects will be otherwise 
healthy older adults with no history of neurological disease or disorder who meet criteria to 
participate in the intervention and do not have any contraindications for MRI. Subjects will be 
randomized to one of the two arms, stratified by APOE4 status.   

 
 
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STUDY TEAM ROSTER 
○ 
PI:  
 
 
Judy Pa, PhD 
 
2025 Zonal Avenue 
 
Los Angeles, CA 90033 
 
Tel: (323) 442-7246 
 
Fax: (323) 442-0137 
 
judypa@usc.edu 
 
Main responsibilities/Key roles: Supervising all aspects of the trial 
○ 
Co-Investigators: 
Peter Conti, MD 
 
Molecular Imaging Center 
 
2250 Alcazar St., CSC 103 
 
Los Angeles, CA 90033 
 
Tel: (323) 442-3858 
 
Fax: (323) 442-3253 
 
pconti@usc.edu 
 
Main responsibilities/Key roles: Incidental findings reports 
Carolee Winstein, PhD, PT, FAPTA 
 
Biokinesiology & Physical Therapy 
 
1540 Alcazar St., CHP 155 
 
Los Angeles, CA 90089 
 
Tel: (323) 442-2903 
 
Fax: (323) 442-1515 
 
winstein@usc.edu 
 
Main responsibilities/Key roles: Clinical trial expertise 
Beth Fisher, PhD, PT, FAPTA 
 
Biokinesiology & Physical Therapy 
 
1540 Alcazar St., CHP 155 
 
Los Angeles, CA 90089 
 
Tel: (323) 442-2796 
 
Fax: (323) 442-1515 
 
bfisher@usc.edu 
 
Main responsibilities/Key roles: Physical function expertise 
Hussein Yassine, MD 
 
Medicine 
 
2250 Alcazar St., CSC 210 
 
Los Angeles, CA 90033 
 
Tel: (323) 442-1909 
 
Fax: (323) 442-4103  

 
 
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hyassine@usc.edu 
 
Main responsibilities/Key roles: APOE, Biomarkers lab 
Wendy Mack, PhD 
 
Preventative Medicine 
 
2001 N. Soto Street, SSB 202Y 
 
Los Angeles, CA 90089 
 
Tel: (323) 442-1820 
 
Fax: (323) 442- 
 
wmack@usc.edu 
 
Main responsibilities/Key roles: Biostatistician of record 
 
 
 
 

 
 
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1 
STUDY OBJECTIVES 
1.1 Primary Objective 
The primary objective is to evaluate change in the outcomes of fMRI brain signal and behavioral 
performance during a mnemonic similarity memory task after a 6-month exercise intervention. 
We hypothesize that the exercise arm will have differences in hippocampal brain signal and 
performance relative to the health education control arm.  
1.2 Secondary Objectives 
Secondary objectives will examine change in outcomes of brain derived neurotrophic factor 
(BDNF), amyloid PET, physical function and aerobic fitness after a 6-month exercise 
intervention. We hypothesize that the exercise arm will have differences in amyloid PET 
accumulation, higher BDNF levels and improved aerobic fitness relative to the health education 
control arm.  
2 
BACKGROUND AND RATIONALE  
2.1 Background on Condition, Disease, or Other Primary Study Focus 
Alzheimer’s disease (AD) is the most common cause of cognitive impairment in older adults and 
affects 36 million people worldwide.1 Results from several large pharmacological trials 
prompted a paradigm shift to initiate treatment earlier in AD. 2-3 Mild cognitive impairment 
(MCI), a preclinical stage of AD, may be an optimal period for initiating treatment. Exercise has 
emerged as an exciting, lifestyle intervention to help remediate cognitive loss or delay onset of 
dementia.8-10 However, to fully capitalize on the investments in exercise and to discover the 
maximum treatment potential of exercise, the brain mechanisms supporting exercise for older 
adults with MCI must be understood.28-30 While a large body of literature has found that exercise 
enhances cognition, very few intervention studies have investigated the brain mechanisms 
responsible for these benefits.31-36 
 
The positive effects of exercise on cognitive function in healthy older adults were first shown 
over a decade ago when aerobic walking improved executive function after 6 months, relative to 
stretching.30 Increases in gray and white matter brain volume after a 6-month aerobic exercise 
intervention were later reported, and this finding was corroborated by a recent 12-month aerobic 
exercise study that showed increased hippocampal volume, improved memory function, and 
increased BDNF in healthy older adults.28,37 Overall, the majority of studies on exercise that 
examined brain volume and cognition were in healthy older adults, leaving exercise effects on 
brain mechanisms in at-risk populations, such as MCI, largely unexplored. In a home-based 
exercise intervention, MCI patients were randomized to an exercise of choice, usually walking, 
or to a usual care control arm. The walking arm improved on the ADAS-Cog (Alzheimer's 
Disease Assessment Scale- cognitive subscale) by 0.26 points while the usual care arm actually 

 
 
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declined by 1.04 points.9-10 Two studies to date, have examined the effects of exercise on brain 
function using fMRI in an intervention study.37 These studies provided a glimpse into brain 
changes in MCI patients after exercise, but there is a need for a comprehensive study to 
understand how impaired brain mechanisms can be remediated with aerobic and strengthening 
exercise to support the cognitive benefits of exercise. 
 
Using advanced brain imaging and key growth factor measures, the proposed study will take a 
cross-disciplinary approach to investigate the effects of exercise on brain mechanisms in 
individuals with MCI. Results from this study will provide critical insight into the brain 
mechanisms that drive the benefits of exercise on cognitive outcomes, will supply evidence of 
the intervention target and engagement, and will give neuroscientific support for public health 
interventions to increase exercise in older adults at risk for AD. 
2.2 Study Rationale 
Using an aerobic walking intervention program, we will conduct an unprecedented investigation 
of brain mechanisms supporting exercise, from large-scale connectivity, and localized function 
and structure of the hippocampus, to lower-level growth factors. This project will recruit 60 
sedentary older adults between 55-80 years of age with early MCI. Based on the Surgeon 
General’s recommendation for moderate-intensity aerobic in older adults, participants will be 
randomly assigned to: aerobic walking for 30 mins, 5 days/week for 6 months OR health 
education non-active control for 6 months, with 30 participants in each arm.14-15 Primary 
outcome measurements include regional brain activity in the hippocampus and behavioral 
performance on the Mnemonic Similarity Task (MST). Additionally, we will examine secondary 
outcomes of brain volume, amyloid burden, cognition and brain derived neurotrophic factor 
(BDNF) associated with exercise. Importantly, all participants will be scanned with amyloid and 
tau PET tracers at baseline and follow-up to examine the contribution of amyloid and tau 
aggregation on brain function and to serve as an AD risk biomarker.  
3 
STUDY DESIGN 
LEARNit is a single-site, single-blind, randomized controlled trial comparing the effects of 
moderate aerobic walking (55-80% target HR) versus healthy living education reading in older 
adults who perform at least 1 standard deviation below normative values for their age (N=60). 
Intervention is performed at home or in the participant’s local community (e.g., home or 
neighborhood). Intervention duration is 6 months. Randomization for this single-blind study was 
achieved using a permuted block schema with block sizes of 2 and 4. Participants were 
randomized in a 1:1 allocation to either aerobic walking or healthy living education arms and 
stratified by APOE4 carrier status (yes/no) based on saliva APOE phenotyping.  

 
 
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4 
SELECTION AND ENROLLMENT OF PARTICIPANTS  
Selected and recruited participants were community-dwelling older adults between the ages of 
55-80 years of age. Target enrichment of 20% underrepresented race/ethnic groups was a 
priority.  
4.1 Inclusion Criteria  
1. Male or female 
2. English speaking 
3. 55-80 years old 
4. Exhibits sedentary behavior (defined as engaging in moderate intensity exercise, such as 
brisk walking for < 60 minutes total per week for the last 3 months) 
5. MMSE score > 26 
6. Perform 1 SD below normative values on neuropsychological tests  
7. Absence of dementia 
8. Ability to complete both fMRI and PET scans 
9. Physical ability to complete exercise program as determined by performance on physical 
screen by certified PT indicating it is safe to participate in the intervention. 
4.2 Exclusion Criteria  
Candidates meeting any of the exclusion criteria at baseline will be excluded from study 
participation and then list each criterion. 
1. Participants meet criteria for dementia (DSM-IV) 
2. History of a neurological disorder 
3. Head trauma with loss of consciousness > 10 minutes 
4. Current psychiatric illness, including depression (GDS>7) 
5. Severe sensory deficits 
6. Current or substantial history of substance abuse 
7. Diabetes 
8. Inability to perform intervention (e.g., inability to walk without assistance)  

 
 
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9. Contraindications to MRI scan (e.g., pacemaker). 
4.3 Study Enrollment Procedures  
Recruitment: 
Participants will be recruited from the community using local media outlets, like the newspaper, 
radio, flyers and local community groups, including senior centers, retirement communities and 
churches. The projected enrollment is 25 participants per year. 
Screening and Eligibility Criteria: 
All study candidates will first be telephone-screened for information on demographics, exercise 
level, medications, and MRI contraindications. If eligible after the telephone screening, 
participants will receive an in-person evaluation including a detailed history, neuropsychological 
screening battery assessing memory and executive function. Screening for depression will be 
done using the self-report 15-item Geriatric Depression Scale. Participants will be deemed 
eligible, meeting criteria for a diagnosis of cognitive decline, when performance on at least one 
measure of the neuropsychological battery is one standard deviation below age, education and 
gender matched norms.  
Screening Records: 
Detailed documentation for all subjects who are screened will be kept in the REDCap database 
including ineligibility documentation.  
Eligibility Criteria: 
A summary of each participant’s in-person screening and eligibility data is included in the 
REDCap database in the Screening Key form. 
Consenting: 
Consent will be obtained from participants at the beginning of the Screening Visit prior to any 
testing. The Project Manager, Coordinator or PI will review the consent form (IFC) with 
participants. One of the collaborating MDs (e.g., Dr. Peter Conti) will be available to answer 
questions (by phone as necessary) and will sign the consent form.  
A copy of the signed consent form will be offered to all participants for their records. The 
original signed consent form will be stored in a locked cabinet in the PI’s office. 
Randomization: 
Randomization for this single-blind study was achieved using a permuted block schema with 
block sizes of 2 and 4. Participants were randomized in a 1:1 allocation to either aerobic walking 
or healthy living education arms and stratified by APOE4 carrier status (yes/no) based on saliva 
APOE phenotyping.  

 
 
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5 
STUDY INTERVENTIONS  
5.1 Interventions, Administration, and Duration  
At-home Walking Program: Participants enrolled in the at-home walking program will have a 
program tailored to them. A trained exercise therapist will assist them in designing this program 
to carry out for a period of 6 months. Over the first 6 weeks subjects will engage in walking 
exercises that gradually increase in duration up to 150 minutes/week. The interventionist will 
work closely with the subject during this period to ensure proper safety, positioning, and 
intensity. From week 6 and on, participants will continue walking at 150 minutes/week. At the 
end of the study, subjects will receive a packet of reading materials for the at-home health 
education program to independently engage in the other intervention program, if they wish. 
At-home Health Education Program: Participants will be asked to participate in an at -home 
health education program for a period of 6 months that consists of a behavioral intervention 
packet of reading materials. Participants will be asked to read about two topics per month from 
the packet. The interventionist will work closely with them during the first 6 weeks to ensure 
understanding of the program and continue to visit them to ensure compliance through the 
duration of the intervention. Following the program, they will be offered the opportunity to 
attend an educational session on exercise with the interventionist and will receive materials on 
engaging in an at home walking program. 
All participants will be asked to wear a physical activity-monitoring device for the intervention 
period of 6 months. 
Participants will be asked to complete surveys, questionnaires, and keep an activity diary to be 
mailed in, collected by the interventionist, completed online or by phone. 
 
5.2 Handling of Study Interventions  
 
Aerobic Walking Arm: 
 
The aerobic group will participate in aerobic walking for a total of 3-5 days/week for 6 months. 
There is a misconception of what constitutes beginner, intermediate and advanced levels of 
aerobics. It is exercise intensity level that determines heart rate.  The target heart rate range for 
participants in the aerobic training group is 60 to 75% of their maximum heart rate. Since all 
participants are 55 years or older, simple walking activities will be all that is required to achieve 
this target heart rate range. All the participants in this study are inactive and may not have much, 
if any, prior aerobic experience. The aerobic walking program is detailed below. Each session 
will include warm-up, aerobic, and cool-down periods.  
 
Components of Aerobic Training Exercise: 
 

 
 
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Warm-up (5 minutes) – Each exercise session will begin with an easy warm-up. Instruct the 
participant to use a heart rate zone of less than 50% of Max Heart Rate to guide your warm-up.  
As a general guideline, a warm-up can be something as simple as walking for 5 minutes at a 
slower pace.   
 
Aerobic Segment (30 minutes) – The objective is to increase subjects’ heart rates to 60% to 75% 
of their maximum. Subjects will measure their heart rate at 15 minutes into the walking period 
and at the end of walking.  At the end of the aerobic training session, have subjects record their 
heart rates and perceived exertion in their Exercise Diaries.  
 
Cool-down (5 min) – Aerobic activity should always be followed by a cool-down period. The 
participant will slow down walking speed in the last 5 minutes of the walking session. Instruct 
the participant to “slow down your movement and try to get your heart rate back down to less 
than 50% of Max Heart Rate. “ Perceived exertion should be checked again to be sure everybody 
is recovering adequately 
 
Progression of Aerobic Exercise: 
 
Ideally participants will progress the walking program over the first 4 weeks with the following 
schedule: weeks 1-2 will consist of 40 minutes for 2 days/week (including 5 minutes warm-up, 
cool-down and 30 minutes of continuous walking). By weeks 3-4 subjects will have added the 
3rd walking day at 40 minutes. Thus, by week 5 up to 6 months, subjects are participating fully 
in the aerobic component of the program (40 minutes total including 5 minutes warm-up, 30 
minutes continuous aerobic walking and 5 minute cool-down for at least 3 days/week). However, 
it is likely that not all subjects in the aerobic group can progress at this rate. In these cases, the 
interventionist will tailor a program of increasing duration per session and increasing frequency 
per week to eventually achieve the complete walking program of 40 minutes.  
 
Structure of aerobic exercise: minimum of 3 days per week (45 minutes each) walking, up to 5 
days per week (30 minutes each).  
 
If a subject has limited endurance and can only manage 10 minutes of continuous aerobic 
exercise during the first session before complaining of fatigue or RPE of ‘very hard exertion,’ 
the following adapted schedule may be used such that the subject is participating fully by Week 
6. 
 
Health Education Arm: 
 
Interventionists will have equivalent contact with participants from the health education to that 
of the aerobic walking group. They will visit weekly for the first four weeks, biweekly at weeks 

 
 
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6 and 8, with a monthly visit thereafter.  Additional bi-weekly phone calls will be made in 
between the monthly visits. The health education protocol will consist of a home-based 
behavioral intervention packet of reading materials and videos for lifestyle practices including 
nutrition, stress management and risk factors for negative health outcomes (e.g., smoking, etc).  
 
At the Orientation Visit (Week 0), subjects will complete the Health Education Contract with the 
interventionist and select their preferred topic order. Participants will be given a binder for the 
reading materials. There will be approximately two topics per month to read and participants will 
answer 4-5 reflection questions per week via electronic journal or on paper, to measure 
adherence. Because providing ‘choices’ to the subjects has been shown to greatly impact 
adherence, the subjects will be given the choice to complete 13 out of 15 of the topics listed 
above. They may order the topics however they wish and select the educational topic they want 
to study for every two week period. Participants should be given the new topics at each home 
visit.  
 
The topic choices for each participant will be recorded in REDCap using the Health Education 
Topic Order instrument. This form must be completed within a week of the Orientation Visit as 
it links the appropriate reflection questions to the reading journals.   
 
Arm Visit Matrix 
Visit 
Aerobic Walking Arm 
Health Education Arm 
Week 0 
Home Visit 
Home Visit 
Week 1 
Home Visit 
Home Visit 
Week 2 
Home Visit 
Home Visit 
Week 3 
Home Visit 
Home Visit 
Week 4 
 
 
Week 5 
Home Visit 
Home Visit 
Week 6 
 
 
Week 7 
Home Visit 
Home Visit 
Week 8 
 
 
Week 9 
Phone Call 
Phone Call 
Week 10 
 
 
Week 11 
Home Visit 
Home Visit 
Week 12 
 
 
Week 13 
Phone Call 
Phone Call 
Week 14 
 
 
Week 15 
Home Visit 
Home Visit 
Week 16 
 
 

 
 
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Week 17 
Phone Call 
Phone Call 
Week 18 
 
 
Week 19 
Home Visit 
Home Visit 
Week 20 
 
 
Week 21 
Phone Call 
Phone Call 
Week 22 
 
 
Week 23 
Home Visit 
Home Visit 
Week 24 
 
 
Week 25 
Phone Call 
Phone Call 
 
6-month Visit 
6-month Visit 
 
5.3 Concomitant Interventions 
5.3.1 Allowed Interventions 
Participants are allowed to continue or begin any medications or treatments prescribed or 
recommended by their physicians. Other studies involving research intervention are not allowed.  
5.3.2 Required Interventions  
None 
5.3.3  Prohibited Interventions 
Other studies involving research intervention are not allowed.  
5.4 Adherence Assessment  
 
 
 
 
 
Adherence will be assessed using several strategies, including Exercise Journal entries, and 
adherence outcomes obtained during the interventionist visits that include completed days of 
activity, exercise duration, time in target HR zone, subjective ratings of perceived effort, and 
achieved HR.  
 
 
 
 
The study team and interventionists will implement behavioral management strategies to support 
a positive exercise environment. The goal in LEARNit will be for participants to complete at 
least 80% of the prescribed exercise sessions. Participants who miss one or more exercise 
sessions each week will be contacted by their interventionist or a study team member to provide 
encouragement and help generate a plan to help the participant complete the training sessions the 
following week. In cases where adherence is an ongoing problem, the study team may meet with 
the interventionist to discuss strategies to increase adherence (e.g., goal setting, reinforcers 
identified and scheduled, etc.). 
Theory and evidence-based approach to increase adherence. A challenge of employing a 
behavioral intervention is achieving program adherence and completion, particularly in 

 
 
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individuals with MCI. To increase the likelihood of meeting these goals, the interventionist will 
draw upon principles of social-cognitive and health belief models for personal and 
environmental factors and perceived barriers and benefits.38-40 The interventionist will begin by 
establishing rapport and trust with the participant by gaining an understanding of the 
participant’s perspective on the intervention and any perceived barriers to increasing self-
efficacy.41-43 This will be followed by an introduction and rationale for the intervention, 
education on the benefits of exercise, and addressing home-life challenges. The program steps of 
each intervention will be detailed in a manual for the interventionist to be performed by the 
interventionist with the participant during two 1-hour sessions during the first 2 weeks.  
To encourage adherence and accountability, these steps will be followed: 1) The interventionist 
will develop a contract with the participant and identify motivating factors for compliance and 
increasing positive health outcomes during the first 2 home visits, and a contract copy will be 
kept in the behavioral modification packet for the participant. 2) The participant will complete an 
exercise journal of activity completion (start/stop time), heart rate during activity, and any 
adverse events. It has been highly successful for collecting confidential information for our 
current projects and will be used for the participants’ journal. 3) The interventionist will conduct 
weekly phone calls for the first 6 weeks and bi-weekly calls thereafter to answer questions, 
reinforce safe practices, and probe any adverse events. Research staff will monitor journals for 
compliance and address any identified concerns. The interventionist will complete monthly 
home visits to provide encouragement and retrieve pedometer data as well as the exercise 
journal. 4) The study coordinator will mail monthly newsletters to the participants on the health 
benefits of exercise (or healthy lifestyles for the control arm). 5) The interventionist will perform 
a fidelity check during 2 of the home visits. Specifically, the interventionist will perform the 
program with the participant (walking, strengthening activities) to observe the participant’s 
completion and use motivational techniques to provide feedback and additional instruction if 
needed. If the participant must travel away from the home during the intervention period of 12 
months, the interventionist will work with the participant to establish a plan for maintaining 
activity during travel time. 6) To mitigate possible reduced motivation over time, we will 
conduct 2 “refresher course” at the midway point of the intervention. The steps of using a 
contract and journal to increase adherence in MCI have been used by Drs. Winstein and Fisher, 
Co-Investigators and are detailed in the Manual of Procedures (MOP). 
 
 
 

 
 
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5.5 
SCHEDULE OF EVALUATIONS 
 
 
Schedule of Evaluations 
  
  
  
  
Screen 
BL 
 6 mo 
Study description / informed consent 
• 
 
 
Neuropsychological Screening 
• 
 
 
Physical Assessment for study inclusion 
• 
 
 
Saliva Collection 
• 
 
 
Accelerometer Tracking 
• 
• 
• 
Modified Physical Function Test 
 
• 
• 
Treadmill walking test 
 
• 
• 
Serum collection 
 
• 
• 
Neuroimaging  
 
• 
• 
   -Task and resting state fMRI 
 
• 
• 
   -T1 anatomical  
 
• 
• 
-Amyloid, Tau PET scan 
 
• 
• 
Neuropsychological Testing 
 
• 
• 
   -Memory, Exec Function 
 
• 
• 
Interventionist home visit (monthly) 
 
• 
• 
Participant phone contact (bi-weekly) 
 
• 
• 
Exit interview 
 
 
• 

 
 
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6 
Description of Evaluations  
6.1 Screening Evaluation 
Visit 1 (Screening Visit): 
• Informed Consent (IFC Review): Review and sign the informed consent form. The 
study investigators will answer any questions participants may have about the study. 
• Evaluation: Subjects will receive an in--person evaluation including a review of 
medical history, physical screen and a comprehensive neuropsychological screening 
battery assessing memory and attention. Subjects will be asked to provide a saliva 
sample. 
30-Day Compliance Test 
• Physical Activity Monitoring: Subjects will be given an accelerometer and asked to 
wear the device at home for a period of 30 days. 
6.1.1 Enrollment, Baseline, and/or Randomization 
Baseline Assessments (2-day Visit): 
The following procedures will take place over 2 days at the baseline visit. Total participation 
during the 2--day period is not anticipated to exceed 10 hours. (For example, baseline 
participation may be 6 hours on day one and 4 hours on day two). 
● Cognitive Tasks: Subjects will be asked to perform one or more memory or attention 
task(s). They may be presented with images, videos, words, numbers, or sounds. The 
cognitive testing session should last no more than two hours. 
● Questionnaires: Subjects will be asked to complete one or more surveys or questionnaires 
on paper or in electronic format. Completing these documents should take no more than 
30 minutes. 
● fMRI Scan: Subjects will be asked to perform cognitive tasks while we use magnetic 
resonance imaging (MRI) to measure changes in activity in the brain. Estimated time in 
the MRI machine will be about 60 to 90 minutes. The entire session should last no more 
than 3.5 hours including said cognitive tasks. 
● PET/CT Scan: Subjects may be asked to participate in 2 positron emission tomography 
(PET/CT) scans per visit (2 at Baseline and 2 at 6-month visit). After a delay of about 75 
minutes to allow the tracers to be absorbed, you will be moved into the PET/CT scanner 
for about 30 minutes. 
● Blood Draw: Subjects will be asked to have a blood draw. This will be a fasting blood 
draw. We will collect about 50 mL of blood. 
● Physical Assessment: Subjects will be asked to complete a physical function test. 
Subjects will be asked to perform a number of everyday activities. The second 
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(CERC) while we monitor heart rate. Subjects will be monitored by trained study staff. 
Finally, body composition and body weight will be measured at the same time on the 
Biospace InBody 520 device. Subjects will stand on the scale-like device while grasping 
two handles, one in each hand. The device works by sending a very low- voltage 
electrical signal through the body to determine water content and body fat percentage. 
The voltage is so low that subjects cannot feel it. The testing will take about 45 minutes. 
● Randomization: Subjects will be assigned randomly, much like tossing a coin, to a study 
group. They will be randomized into one of two health programs, and will have a 50% 
chance of being placed in either group. One group will participate in an at-home walking 
program and the other an at-home health education program. 
 
6.1.2 Follow-up Visits / Completion/Final Evaluation 
Follow-up 6-month Visit (2-day Visit): 
The following procedures will take place over 2 days at the follow-up visit. Total participation 
during the 2-day period is not anticipated to exceed 10 hours. (For example, participation may be 
6 hours on day one and 4 hours on day two). 
● Cognitive Tasks: Subjects will be asked to perform one or more memory or attention 
task(s). They may be presented with images, videos, words, numbers, or sounds. The 
cognitive testing session should last no more than two hours. 
● Questionnaires: Subjects will be asked to complete one or more surveys or questionnaires 
on paper or in electronic format. Completing these documents should take no more than 
30 minutes. 
● fMRI Scan: Subjects will be asked to perform cognitive tasks while we use magnetic 
resonance imaging (MRI) to measure changes in activity in the brain. Estimated time in 
the MRI machine will be about 60 to 90 minutes. The entire session should last no more 
than 3.5 hours including said cognitive tasks. 
● PET/CT Scan: Subjects may be asked to participate in 2 positron emission tomography 
(PET/CT) scans per visit (2 at Baseline and 2 at 6-month visit). After a delay of about 75 
minutes to allow the tracers to be absorbed, you will be moved into the PET/CT scanner 
for about 30 minutes. 
● Blood Draw: Subjects will be asked to have a blood draw. This will be a fasting blood 
draw. We will collect about 50 mL of blood. 
● Physical Assessment: Subjects will be asked to complete a physical function test. 
Subjects will be asked to perform a number of everyday activities. The second 
assessment is a treadmill walking test performed at the clinical exercise research center 
(CERC) while we monitor heart rate. Subjects will be monitored by trained study staff. 
Finally, body composition and body weight will be measured at the same time on the 
Biospace InBody 520 device. Subjects will stand on the scale-like device while grasping 

 
 
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two handles, one in each hand. The device works by sending a very low- voltage 
electrical signal through the body to determine water content and body fat percentage. 
The voltage is so low that subjects cannot feel it. The testing will take about 45 minutes. 
7 
SAFETY ASSESSMENTS  
Adverse events are recorded via electronic recording directly into REDCap by the 
participant in the form of “Have there been any changes in your health status?” Any 
reporting of an AE triggers an alert to the study team in the REDCap system. 
7.1 Adverse Events and Serious Adverse Events  
 
An adverse event (AE) is generally defined as any unfavorable and unintended diagnosis, 
symptom, sign (including an abnormal laboratory finding), syndrome or disease which either 
occurs during the study, having been absent at baseline, or if present at baseline, appears to 
worsen. Adverse events are to be recording regardless of their relationship to the study 
intervention. 
 
A serious adverse event (SAE) is generally defined as any untoward medical occurrence that 
results in death, is life threatening, requires inpatient hospitalization or prolongation of existing 
hospitalization, results in persistent or significant disability/incapacity, or is a congenital 
anomaly. 
 
7.1.1  Evaluation of Safety Measures 
 
Event and participant count of the following will be summarized overall and by treatment group 
for the entire cohort: 
 
• AE: Overall and by MedDRA System Organ Class 
• AE: MedDRA Preferred Term 
• SAE: Overall and by MedDRA System Organ Class 
• SAE: MedDRA Preferred Term 
• SAE Definitely Related to Intervention: Overall and by System Organ Class 
• Hospitalization 
• Deaths 
 
Comparisons of the number of participants with at least one AE, SAE, SAE definitely related to 
Intervention and Death will be examined between Intervention groups using the Fisher's Exact 
test. 

 
 
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7.2 Reporting Procedures 
AEs will be reported to the IRB according to the timing specifications laid out by the governing 
body. SAEs will be reported within 24 hrs of reporting to the Safety Monitor and the IRB. 
7.3 Follow-up for Adverse Events 
The follow-up of adverse events occurs while the participant is actively engaged in the study 
through electronic inquiry. 
7.4 Safety Monitoring 
This trial was monitored by an NIH-approved Data and Safety Monitoring Officer, Dr. Sharon 
Sha, MD, Chief of Memory Disorders Division at Stanford University.  
 
Sharon J. Sha, MD, MS 
Clinical Professor 
Chief, Memory Disorders Division 
Associate Vice Chair, Clinical Research 
Neurology and Neurological Sciences 
Stanford Center for Memory Disorders 
Director, Huntington’s Disease Center of Excellence and Ataxia Clinic 
Co-Director, Lewy Body Dementia Research Center of Excellence 
  
Stanford Neuroscience Health Center 
453 Quarry Road, MC 5979 
Palo Alto, CA 94304 
ssha1@stanford.edu 
memoryresearch@stanford.edu 
http://med.stanford.edu/neurology.html 
 
8 
INTERVENTION DISCONTINUATION   
 
Investigators will make every effort to maximize participant retention. If a participant expresses 
a desire to stop the intervention, the study team will meet to discuss the participant’s existing 
challenges and possible approaches to reduce burden to encourage continued participation if at 
all possible (e.g., adapt exercise routine to reduce pain or increase interest, problem-solve 
regarding adherence issues, etc.). 
If a participant continues to decline further participation, or if an investigator discontinues the 
intervention, an exit interview should be completed as soon as possible immediately following 
intervention discontinuation.  

 
 
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All early intervention discontinuation participants will be strongly encouraged to complete the 
month 6 assessment visit.  
 
 
 
 
 
 
 
 
 
Reasons for Early Discontinuation  
 
 
 
 
 
Participants are allowed to voluntarily discontinue participation in this trial at any time. Due to 
the low safety risk of this intervention, we do not anticipate early discontinuation due to safety 
concerns. 
 
9 
STATISTICAL CONSIDERATIONS  
9.1 General Design Issues  
Hypotheses regarding the primary cognitive outcome, and secondary and exploratory outcomes 
will be tested using a linear mixed model for repeated measures (MMRM) including indicator 
variables for time, intervention, time-by-intervention interaction term, and additional covariates 
meeting confounding criteria below. A participant-level random intercept will be specified; 
significance of random slopes will be tested and included in the model if significant. Age and 
sex/gender will be treated as potential confounders. Potential confounders will be identified from 
randomization imbalance and based on their effect on the magnitude of treatment. A variable 
will be treated as confounder if adding it to the model changes the value of treatment estimate by 
more than 15%. APOE4 will be included in all models because it was a randomization 
stratification factor. For fMRI hippocampal BOLD analysis, within subject changes in activity 
will be calculated for each individual subject in those with both baseline and follow-up fMRI 
data. Subject data will be normalized to group template space which is comprised of timepoint 1 
and 2 to reduce measurement error or individual subject bias. Analyses will be performed by 
staff blinded to the treatment group. 
9.2 Sample Size and Randomization 
 
Given the planned sample size of 30 participants per group, the magnitude of effect observable 
for this analysis are described below:  
This study was powered based on the fMRI primary outcome. Group sample sizes of 30 achieve 
80% power for each test to detect an effect size of at least 0.736 and a type I error of 0.05 using a 
two-sided two-sample equal-variance t-test. There are 2 primary outcomes (cognition, imaging), 
but because they are very different measures, the alpha level will remain at 0.05 for each primary 
outcome test. There will be no alpha-level correction for secondary or exploratory outcomes. 
Preliminary data provide an estimated standard deviation of 0.25 z-score units for fMRI network 

 
 
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function, indicating that we will be able to detect differences of 0.184 z-score units in brain 
function between groups. The effect sizes for fMRI BOLD activity and network function 
reported in previous physical activity intervention studies in older adults ranged from 1.17-1.24, 
thus we have high power to detect significant differences in brain function.  
9.2.1 Treatment Assignment Procedures 
 
 
 
We used a randomized, single-blind study design where randomization was achieved using a 
permuted block schema with block sizes of 2 and 4. Participants were randomized in a 1:1 
allocation to either aerobic walking or healthy living education arms and stratified by APOE4 
carrier status (yes/no) based on saliva APOE phenotyping. Research suggests that APOE4 status 
may affect response to exercise, thus carriers were balanced across groups.  
 
 
 
9.3 Interim analyses and Stopping Rules 
No interim analyses or stopping rules were applied for this clinical trial. 
9.4 Outcomes  
Hypotheses regarding the primary cognitive outcome, and secondary and exploratory outcomes 
will be tested using a linear mixed model for repeated measures (MMRM) including indicator 
variables for time, intervention, time-by-intervention interaction term, and additional covariates 
meeting confounding criteria below. A participant-level random intercept will be specified; 
significance of random slopes will be tested and included in the model if significant. Age and 
sex/gender will be treated as potential confounders. Potential confounders will be identified from 
randomization imbalance and based on their effect on the magnitude of treatment. A variable 
will be treated as confounder if adding it to the model changes the value of treatment estimate by 
more than 15%. APOE4 will be included in all models because it was a randomization 
stratification factor. 
 
For fMRI hippocampal BOLD analysis, within subject changes in activity will be calculated for 
each individual subject in those with both baseline and follow-up fMRI data. Subject data will be 
normalized to group template space which is comprised of timepoint 1 and 2 to reduce 
measurement error or individual subject bias. Analyses will be performed by staff blinded to the 
treatment group. 
9.4.1 Primary outcome   
The primary outcome measures will be collected during fMRI acquisition at baseline and 6-
month follow-up. Participants will complete the mnemonic similarity test (MST) during the 
fMRI scan to provide both primary outcome measures of hippocampal BOLD signal and MST 
performance, which is assessed using the Lure Discrimination Index.  
 

 
 
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● Hippocampal BOLD signal: change in hippocampal BOLD signal during the MST task 
defined as differences in activity between baseline and 6-month as the outcome measure. 
Change can be in either direction. 
 
● Lure Discrimination Index (LDI): change in the LDI score, defined as 6-month – 
baseline, as the outcome measure will be analyzed using the MMRM approach. Higher 
change scores indicate improvement in LDI performance. 
 
9.4.2 Secondary outcomes   
Secondary analyses will be conducted using the same MMRM statistical approach described 
above for the analysis of all secondary outcomes. 
 
• Single-stage treadmill test: change in the VO2 max score from the single-stage treadmill 
test defined as 6-month – baseline as the outcome measure. Higher change scores indicate 
improvement. 
• Physical function test score: change in the Physical Function test score defined as 6- 
month – baseline as the outcome measure. Higher change scores indicate improvement. 
• Brain-derived neurotrophic factor (BDNF) level: change in total plasma BDNF defined 
as 6-month – baseline level as the outcome measure. Higher change scores are better. 
• Amyloid PET SUVR: change in global amyloid composite score including the mean of 
frontal, temporal, parietal cortex SUVRs defined as 6-months – baseline will be the 
outcome measure. Lower change scores are better. 
9.5 Data Analyses 
Hypotheses regarding the primary cognitive outcome, and secondary and exploratory outcomes 
will be tested using a linear mixed model for repeated measures (MMRM) including indicator 
variables for time, intervention, time-by-intervention interaction term, and additional covariates 
meeting confounding criteria below. A participant-level random intercept will be specified; 
significance of random slopes will be tested and included in the model if significant. Age and 
sex/gender will be treated as potential confounders. Potential confounders will be identified from 
randomization imbalance and based on their effect on the magnitude of treatment. A variable 
will be treated as confounder if adding it to the model changes the value of treatment estimate by 
more than 15%. APOE4 will be included in all models because it was a randomization 
stratification factor. 
 
For fMRI hippocampal BOLD analysis, within subject changes in activity will be calculated for 
each individual subject in those with both baseline and follow-up fMRI data. Subject data will be 
normalized to group template space which is comprised of timepoint 1 and 2 to reduce 
measurement error or individual subject bias. 

 
 
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10 
DATA COLLECTION AND QUALITY ASSURANCE 
10.1 Data Collection Forms 
Data is collected using REDCap forms. Outcome assessments are collected by blinded assessors 
from study staff, the USC Memory and Aging Center, the Clinical Exercise Research Center and 
the USC PET Imaging Center. Subject data is collected using only their study ID, for which the 
key is kept separate and inaccessible by the blinded assessors. Physical copies of the IFC will be 
kept separately from all other study forms in a locked file cabinet, within in a secure office.  
 
Data acquired during the intervention period for completion of intervention tasks was entered 
remotely via REDCap with assistance from the interventionists, as the intervention was home-
based.  
10.2 Data Management  
This was a single site randomized clinical trial. Screening, Baseline and Follow-up visit data was 
collected on site at USC, by the study team, the USC Memory and Aging Center, as well as the 
Clinical Exercise Research Center and the USC PET imaging center. Data was scored when 
needed by the assessor collecting the outcome measures. Imaging data was acquired and 
transferred to the USC NIIN Computer cluster. Clinical report forms were used on paper or 
electronic entry. All paper data were entered into the online digital database, REDCap. REDCap 
data forms were created with built-in value controls and double entry from independent data 
managers was used to ensure accuracy.  
10.3 Quality Assurance  
10.3.1 Training 
Data entry staff were trained on all REDCap forms and cognitive tests as needed. Accuracy of 
data entry was ensured via double entry from 2 independent data managers familiar with the 
forms. Double data entry was used for all paper to electronic transfer scoring. Any discrepancies 
were discussed by the quality control group described below.  
10.3.2 Quality Control Committee  
A committee of study staff was formed to review the cognitive testing and physical function tests 
for consistency and accuracy of scoring. The group met at three separate time frames to review 
data together. Any discrepancies were reviewed and adjudicated by the group for consensus on 
accuracy and scoring values.  

 
 
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10.3.3 Metrics 
REDCap data forms were created with built-in value controls and double entry from independent 
data managers was used to ensure accuracy.  
10.3.4 Protocol Deviations 
Protocol deviations will be captured in REDCap using a protocol deviation form to record details 
and timeline of the deviations. Deviations will be reviewed in real time by the study team to 
determine appropriate action.  
10.3.5 Monitoring 
Consent forms were collected during the initial screening visit, signed by the PI and uploaded to 
the clinical trials portal within 2-4 weeks, before the baseline assessment visit. Outside review 
was conducted by the University to ensure completion at the baseline visit. Participants 
submitted weekly activity reports to the study team using REDCap. Reports were monitored 
weekly and contact was made by the study team or interventionist to ensure completion and 
accuracy.  
 
11 
PARTICIPANT RIGHTS AND CONFIDENTIALITY  
11.1 Institutional Review Board (IRB) Review  
This protocol and the informed consent document (Appendix I) and any subsequent 
modifications will be reviewed and approved by the IRB.  
11.2 Informed Consent Forms 
A signed consent form will be obtained from each participant.   Participants who cannot consent 
for themselves will be excluded from this study. The consent form will describe the purpose of 
the study, the procedures to be followed, and the risks and benefits of participation. A copy will 
be given to each participant. 
11.3 Participant Confidentiality  
Any data, specimens, forms, reports, video recordings, and other records that leave the site will 
be identified only by a participant identification number (Participant ID, PID) to maintain 
confidentiality.  All records will be kept in a locked file cabinet.  All computer entry and 
networking programs will be done using PIDs only. Information will not be released without 
written permission of the participant, except as necessary for monitoring by IRB, the FDA, the 
NIA, and the OHRP. Any data with subject identifiers will be kept in a separate locked file 
cabinet from the Participant Study ID.  

 
 
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11.4 Study Discontinuation  
The study may be discontinued at any time by the IRB, the NIA, the OHRP, the FDA, or other 
government agencies as part of their duties to ensure that research participants are protected. 
 
 
12 
ETHICAL CONSIDERATIONS 
 
  
 
 
 
 
 
Good Clinical Practice  
This study will be conducted in accordance with Good Clinical Practice (GCP) guidelines, as 
defined by the International Conference on Harmonisation (ICH) Guideline, Topic E6, the 
United States Code of Federal Regulations, Title 21, Part 50 (21CFR50) – Protection of Human 
Subjects and Part 56 – IRBs, HIPAA, State and Federal regulations and all other applicable local 
regulatory requirements and laws. 
 
Study personnel involved in conducting this study will be qualified by education, training and 
experience to perform their respective task(s) in accordance with GCP. 
 
No study document shall be destroyed without prior written agreement from the investigator. 
Should the investigator wish to assign study records to another party or move them to another 
location, he/she may do so only with the prior written consent from USC regulatory bodies. 
 
Participant Confidentiality | HIPAA 
 
 
 
 
 
Information about study participants will be kept confidential and managed according to the 
requirements of HIPAA. HIPAA regulations require a signed HIPAA Authorization informing 
the participant of the following: 
 
 
 
 
● What protected health information (PHI) will be collected from participants in this study 
  
 
 
 
 
 
 
● Who will have access to that information and why 
  
 
 
 
 
 
 
● Who will use or disclose that information 
  
 
 
 
 
 
 
● The rights of a research participant to revoke their authorization for use of their PHI 
  
 
 
 
 
 
 
 
In the event that a participant revokes authorization to collect or use PHI, the investigator, by 
regulation, retains the ability to use all information collected prior to the revocation of participant 
authorization. The PI, under the guidance of the local IRB, is responsible for ensuring that all 
applicable HIPAA regulations and State laws are met.  

 
 
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Storage of Biospecimen Samples 
  
 
 
 
 
 
 
 
All biospecimens banked for future AD biomarker research will be stored in freezers at USC. 
Sample tubes are bar-coded and linked to PID only and banked without personal identifiers. The 
presence of the sample is recorded into a computerized inventory database that is managed in 
RedCap and is encrypted and password-protected. 
  
 
 
 
 
 
 
 
 
 
 
 
 
MRI Data Storage 
MRI scans will be labeled with PID. All imaging data will be de-identified using a separate 8 
digit identifier to maintain blinding and are checked by the imaging staff to confirm the absence 
of participant identifying information.  
 
 
 
 
13 
PUBLICATION OF RESEARCH FINDINGS 
Publication of the results of this trial will be governed by the policies and procedures set forth by 
the sponsor, the National Institutes of Health - National Institute on Aging.  
 
  
 
 
  
 
 
 
 
 
 
 

 
 
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REFERENCES 
1. 
Alzheimer's Association. 2012 Alzheimer's disease facts and figures. Alzheimers Dement 
2012; 8(2): 131-68. 
 
2. 
Sperling RA, Jack CR, Jr., Aisen PS. Testing the right target and right drug at the right 
stage. Science translational medicine 2011; 3(111): 111cm33. 
 
3. 
Vellas B, Hampel H, Rouge-Bugat ME, Grundman M, Andrieu S, Abu-Shakra S, 
Bateman R, Berman R, Black R, Carrillo M, Donohue M, Mintun M, Morris J, Petersen R, 
Thomas RG, Suhy J, Schneider L, Seely L, Tariot P, Touchon J, Weiner M, Sampaio C, 
Aisen P. Alzheimer's disease therapeutic trials: EU/US Task Force report on recruitment, 
retention, and methodology. J Nutr Health Aging 2012; 16(4): 339-45. 
 
4. 
Golde TE, Schneider LS, Koo EH. Anti-abeta therapeutics in Alzheimer's disease: the 
need for a paradigm shift. Neuron 2011; 69(2): 203-13. 
 
5. 
Barnes DE, Yaffe K. The projected effect of risk factor reduction on Alzheimer's disease 
prevalence. Lancet Neurol 2011; 10(9): 819-28. 
 
6. 
Middleton LE, Yaffe K. Targets for the prevention of dementia. J Alzheimers Dis 2010; 
20(3): 915-24. 
 
7. 
Middleton LE, Yaffe K. Promising strategies for the prevention of dementia. Arch Neurol 
2009; 66(10): 1210-5. 
 
8. 
Larson EB, Wang L, Bowen JD, McCormick WC, Teri L, Crane P, Kukull W. Exercise is 
associated with reduced risk for incident dementia among persons 65 years of age and 
older. Ann Intern Med 2006; 144(2): 73-81. 
 
9. 
Lautenschlager NT, Cox K, Kurz AF. Physical activity and mild cognitive impairment and 
Alzheimer's disease. Curr Neurol Neurosci Rep 2010; 10(5): 352-8. 
 
10. 
Lautenschlager NT, Cox KL, Flicker L, Foster JK, van Bockxmeer FM, Xiao J, Greenop 
KR, Almeida OP. Effect of physical activity on cognitive function in older adults at risk 
for Alzheimer disease: a randomized trial. JAMA 2008; 300(9): 1027-37. 
 
11. 
Voss MW, Nagamatsu LS, Liu-Ambrose T, Kramer AF. Exercise, brain, and cognition 
across the life span. J Appl Physiol (1985) 2011; 111(5): 1505-13. 
 
12. 
Voss MW, Erickson KI, Prakash RS, Chaddock L, Malkowski E, Alves H, Kim JS, Morris 
KS, White SM, Wojcicki TR, Hu L, Szabo A, Klamm E, McAuley E, Kramer AF. 
Functional connectivity: a source of variance in the association between cardiorespiratory 
fitness and cognition? Neuropsychologia 2010; 48(5): 1394-406. 
 
13. 
Colcombe S, Kramer AF. Fitness effects on the cognitive function of older adults: a meta-

 
 
LEARNit Protocol 
Version 3.0 
April 26, 2024 
 
 
31 
analytic study. Psychol Sci 2003; 14(2): 125-30. 
 
14. 
Physical Activity Guidelines Advisory Committee Report. In: Services. UDoHaH, editor. 
Washington DC; 2008. 
 
15. 
US Department of Health and Human Services CfDCaP. A Report of the Surgeon 
General: Physical Activity and Health for Older Adults. In: Promotion NCfCDPaH, editor. 
www.cdc.gov; 2014. 
 
16. 
Yassa MA, Lacy JW, Stark SM, Albert MS, Gallagher M, Stark CE. Pattern separation 
deficits associated with increased hippocampal CA3 and dentate gyrus activity in 
nondemented older adults. Hippocampus 2011; 21(9): 968-79. 
 
17. 
Brock Kirwan C, Hartshorn A, Stark SM, Goodrich-Hunsaker NJ, Hopkins RO, Stark CE. 
Pattern separation deficits following damage to the hippocampus. Neuropsychologia 2012; 
50(10): 2408-14. 
18. 
Yamamoto H, Gurney ME. Human platelets contain brain-derived neurotrophic factor. 
The Journal of neuroscience : the official journal of the Society for Neuroscience 1990; 
10(11): 3469-78. 
 
19. 
Ebbeling CB, Ward A, Puleo EM, Widrick J, Rippe JM. Development of a single-stage 
submaximal treadmill walking test. Medicine and science in sports and exercise 1991; 
23(8): 966-73. 
 
20. 
Adlard PA, Perreau VM, Pop V, Cotman CW. Voluntary exercise decreases amyloid load 
in a transgenic model of Alzheimer's disease. The Journal of neuroscience : the official 
journal of the Society for Neuroscience 2005; 25(17): 4217-21. Widrick J, Ward A. 
21. 
Ebbeling C, Clemente E, Rippe JM. Treadmill validation of an over-ground walking test 
to predict peak oxygen consumption. European journal of applied physiology and 
occupational physiology 1992; 64(4): 304-8. 
 
22. 
Zanto TP, Pa J, Gazzaley A. Reliability measures of functional magnetic resonance 
imaging in a longitudinal evaluation of mild cognitive impairment. NeuroImage 2014; 84: 
443-52. PMCID: PMC3855402 
 
23. 
Atri A, O'Brien JL, Sreenivasan A, Rastegar S, Salisbury S, DeLuca AN, O'Keefe KM, 
LaViolette PS, Rentz DM, Locascio JJ, Sperling RA. Test-retest reliability of memory 
task functional magnetic resonance imaging in Alzheimer disease clinical trials. Arch 
Neurol 2011; 68(5): 599-606. 
 
24. 
Caceres A, Hall DL, Zelaya FO, Williams SC, Mehta MA. Measuring fMRI reliability 
with the intra-class correlation coefficient. NeuroImage 2009; 45(3): 758-68. 
 
25. 
Bondi MW, Houston WS, Eyler LT, Brown GG. fMRI evidence of compensatory 
mechanisms in older adults at genetic risk for Alzheimer disease. Neurology 2005; 64(3): 
501-8. 

 
 
LEARNit Protocol 
Version 3.0 
April 26, 2024 
 
 
32 
 
26. 
Dickerson BC, Salat DH, Greve DN, Chua EF, Rand-Giovannetti E, Rentz DM, Bertram 
L, Mullin K, Tanzi RE, Blacker D, Albert MS, Sperling RA. Increased hippocampal 
activation in mild cognitive impairment compared to normal aging and AD. Neurology 
2005; 65(3): 404-11. 
 
27. 
O'Brien JL, O'Keefe KM, LaViolette PS, DeLuca AN, Blacker D, Dickerson BC, Sperling 
RA. Longitudinal fMRI in elderly reveals loss of hippocampal activation with clinical 
decline. Neurology 2010; 74(24): 1969-76. 
 
28. 
Brown BM, Peiffer JJ, Martins RN. Multiple effects of physical activity on molecular and 
cognitive signs of brain aging: can exercise slow neurodegeneration and delay Alzheimer's 
disease? Mol Psychiatry 2012. 
 
29. 
Pereira AC, Huddleston DE, Brickman AM, Sosunov AA, Hen R, McKhann GM, Sloan 
R, Gage FH, Brown TR, Small SA. An in vivo correlate of exercise-induced neurogenesis 
in the adult dentate gyrus. Proceedings of the National Academy of Sciences of the United 
States of America 2007; 104(13): 5638-43. 
 
30. 
Erickson KI, Voss MW, Prakash RS, Basak C, Szabo A, Chaddock L, Kim JS, Heo S, 
Alves H, White SM, Wojcicki TR, Mailey E, Vieira VJ, Martin SA, Pence BD, Woods 
JA, McAuley E, Kramer AF. Exercise training increases size of hippocampus and 
improves memory. Proceedings of the National Academy of Sciences of the United States 
of America 2011; 108(7): 3017-22. 
 
31. 
Pa J, Goodson W, Bloch A, King AC, Yaffe K, Barnes DE. Effect of exercise and 
cognitive activity on self-reported sleep quality in community-dwelling older adults with 
cognitive complaints: a randomized controlled trial. Journal of the American Geriatrics 
Society 2014; 62(12): 2319-26. NIHMSID: 654597 
 
32. 
Cox KL, Flicker L, Almeida OP, Xiao J, Greenop KR, Hendriks J, Phillips M, 
Lautenschlager NT. The FABS trial: a randomised control trial of the effects of a 6-month 
physical activity intervention on adherence and long-term physical activity and self-
efficacy in older adults with memory complaints. Prev Med 2013; 57(6): 824-30. 
 
33. 
Barnes DE, Santos-Modesitt, W., Poelke, G., Kramer, A., Castro, C., Middleton, L.E., & 
Yaffe, K. The Mental Activity and eXercise (MAX) Trial: A Randomized, Controlled 
Trial to Enhance Cognitive Function in Older Adults. Arch Intern Med In press. 
 
34. 
Baker LD, Frank LL, Foster-Schubert K, Green PS, Wilkinson CW, McTiernan A, 
Plymate SR, Fishel MA, Watson GS, Cholerton BA, Duncan GE, Mehta PD, Craft S. 
Effects of aerobic exercise on mild cognitive impairment: a controlled trial. Arch Neurol 
2010; 67(1): 71-9. 
 
35. 
Teri L, Logsdon RG, McCurry SM. Exercise interventions for dementia and cognitive 
impairment: the Seattle Protocols. J Nutr Health Aging 2008; 12(6): 391-4. 

 
 
LEARNit Protocol 
Version 3.0 
April 26, 2024 
 
 
33 
 
36. 
Heyn P, Abreu BC, Ottenbacher KJ. The effects of exercise training on elderly persons 
with cognitive impairment and dementia: a meta-analysis. Arch Phys Med Rehabil 2004; 
85(10): 1694-704. 
 
37. 
Herrmann SD, Martin LE, Breslin FJ, Honas JJ, Willis EA, Lepping RJ, Gibson CA, 
Befort CA, Lambourne K, Burns JM, Smith BK, Sullivan DK, Washburn RA, Yeh HW, 
Donnelly JE, Savage CR. Neuroimaging studies of factors related to exercise: Rationale 
and design of a 9month trial. Contemp Clin Trials 2014; 37(1): 58-68. 
 
38. 
Umstattd MR, Hallam J. Older adults' exercise behavior: roles of selected constructs of 
social-cognitive theory. J Aging Phys Act 2007; 15(2): 206-18. 
 
39. 
Lucidi F, Grano C, Barbaranelli C, Violani C. Social-cognitive determinants of physical 
activity attendance in older adults. J Aging Phys Act 2006; 14(3): 344-59. 
 
40. 
Carpenter CJ. A meta-analysis of the effectiveness of health belief model variables in 
predicting behavior. Health Commun; 25(8): 661-9. 
 
41. 
Bandura A. The anatomy of stages of change. Am J Health Promot 1997; 12(1): 8-10. 
 
42. 
Schwarzer R, Lippke S, Luszczynska A. Mechanisms of health behavior change in 
persons with chronic illness or disability: the Health Action Process Approach (HAPA). 
Rehabil Psychol; 56(3): 161-70. 
 
43. 
Jacobs AD, Ammerman AS, Ennett ST, Campbell MK, Tawney KW, Aytur SA, Marshall 
SW, Will JC, Rosamond WD. Effects of a tailored follow-up intervention on health 
behaviors, beliefs, and attitudes. J Womens Health (Larchmt) 2004; 13(5): 557-68. 
 
 
 
 
 
 
 

 
 
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SUPPLEMENT / APPENDICES 
 
Appendix I - Informed Consent Form 
 
Appendix II - Covid - 19 Required Safety Protocols 
 
 
 

 
 
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Appendix I – LEARNit Informed Consent
 

 
 
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Appendix II – COVID-19 Required Safety Protocols 
  
Research staff (Interventionist) practices in the field 
  
o 
Prior to engaging in research activities in any community/field settings, research staff must 
be tested and confirmed negative for COVID-19 in accordance with local testing guidelines 
/ requirements. 
 
o 
Research staff must provide attestation in their USC istar account that they have reviewed 
and understand the plan for completing a daily log of contacts with whom they have had 
direct in-person interaction, including research participants and staff. 
 
o 
Before traveling to community/field research site(s) each day, all research staff must 
complete the Trojan Check university health screening protocol that is required of persons 
when requesting access to USC campuses and buildings.  
o 
https://trojancheck.usc.edu/login 
 
o 
In traveling to/from local community/field research sites, researchers must wear face 
coverings, ensure no more than two people occupancy per vehicle, and/or that local public 
health guidelines are followed when mass transit (bus/metro) is used. Researchers must 
wash hands (or use hand sanitizers) immediately upon departing the vehicle.  High touch 
areas (e.g., keys, door handles, steering wheels, seat belt buckle) must be disinfected before 
and after the field day. 
 
o 
Research staff must enter the community/field setting each day prepared with a sufficient 
number of face coverings and hand sanitizer for all research participants. 
 
o 
Researchers at each off-campus study setting must complete a daily log of contacts with 
whom they have had direct in-person interaction, including research participants and staff, 
for purposes of participant notification.  Contact information must be collected for the 
purpose of participant notification in all of the following cases:   1) any interactions 
occurring at fewer than 6 feet of distance and for more than a period of briefly passing by 
someone, whether indoors or outdoors; and 2) interactions occurring indoors for more than a 
period of 10 minutes.  These rules apply regardless of whether PPE / facial coverings are 
worn during the interaction. 
 
o 
Hand washing is required of research staff every 30 minutes and at the beginning and end of 
each visit to a community or field site.  When hand-washing is not possible, hand sanitizer 
may be used. 
 
o 
If a research staff member feels ill at any point while in the community/field setting, 
he/she/they must discontinue data collection and other research activities immediately, 

 
 
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report illness to the field supervisor and the USC health office, and self-quarantine. 
 
o 
If a research staff member is diagnosed with COVID-19, they must contact USC Student 
Health to discuss and be prepared to share participant notification data with Student Health. 
 
o 
Community/field research activities including data collection should be conducted in 
sparsely populated, low-density spaces and settings – preferably outdoors if at all possible. 
 
o 
Research staff must ensure that research participants and prospective participants in field 
and community settings are assembled and/or seated in a manner that adheres to COVID-19 
safety and risk mitigation standards, including a minimum of 6 feet of physical distancing. 
 
o 
Research staff must provide to each participant an information sheet that explains the 
collection of data for the purpose of participant notification in the case of infection. 
 
o 
Disinfectant wipes must be used in community/field settings to clean pens, clipboards, 
tablets, laptops, and any other equipment or devices immediately before and after sharing 
among research staff or between participants, or every 30 minutes, whichever is more 
frequent.  Hand washing or sanitizing must also be done at these times. 
 
o 
Disinfectant wipes must be used to clean cases, bags, or other containers immediately before 
they are provided to the participant and after they are returned, or every 30 minutes, 
whichever is more frequent.  Hand washing or hand sanitizing must also be done at these 
times.  
 
o 
Physical incentive items (e.g., gift cards) must first be cleaned using disinfectant wipes. 
 
o 
Research participants who do not have a facial covering should be provided with masks 
(paper or cloth as directed by current public health guidelines) and hand sanitizer by the 
research staff  prior to engaging in any research activity including study screening. 
 
o 
Research participants who refuse to wear facial coverings or practice risk mitigation steps 
necessary to protect themselves and others will not be allowed to participate in USC 
research.  (Exceptions for wearing masks shall be granted for children under two years of 
age and persons with disabilities, as recommended by current public health guidelines.) 
 
o 
Research participants must be provided with information about COVID-19 risk mitigation 
and safety practices (i.e., information sheet available through OPRS). 
 
o 
Research participants should be provided information on when and how to access health 

 
 
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care services (http://www.publichealth.lacounty.gov/media/Coronavirus/FAQ.pdf), and a 
list of available social services and resources such as food and domestic violence assistance 
(e.g.,  https://www.211la.org/   and   https://www.1degree.org/). 
 
o 
As with all aspects of community/field research, research participants’ culture, language, 
and health literacy should be considered when communicating COVID-19 safety and risk 
mitigation information and practices.