Functional Studies of Novel Genes Mutated in Primary Ciliary Dyskinesia II: Genotype to Phenotype
Phase: EARLY_PHASE1
Conditions: Primary Ciliary Dyskinesia
Sponsor: University of North Carolina, Chapel Hill

 
Study Title: Functional Studies of Novel Genes Mutated in Primary 
Ciliary Dyskinesia II: Genotype to Phenotype 
 
 
 
NCT number NCT04901715 
Document Date 02/08/2023 
 

Study Title: Functional Studies of Novel Genes Mutated in Primary Ciliary Dyskinesia II: Genotype to 
Phenotype  
 
UNC Principal Investigator (PI): Lawrence E. Ostrowski 
UNC PI Department/Division: Pediatrics/Pulmonary 
 
Number of Sites: 1 
Lead Site: Meadowmont/Eastowne 
Lead Site Principal Investigator: William Bennett, PhD 
Additional Site: USEPA Human Studies Facility 
 
UNC Co-Investigator(s) and Department/Division    
William Bennett, PhD; Medicine/Pulmonary 
Margaret Leigh, MD; Pediatrics/Pulmonary 
Michael Knowles, MD; Medicine/Pulmonary 
Sara Abu-Nasser, DO; Pediatrics/Pulmonary 
Peadar Noone, MD; Medicine/Pulmonary 
 
Study Statistician  
Agathe Ceppe 
Marsico Hall Room 2204,  
125 Mason Farm Road    
Chapel Hill, NC 27599      
 
Sponsor(s) N/A 
 
Source(s) of Funding : NIH; NHLBI 
 
 
 
 
 
 
 
 

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I HAVE READ AND APPROVE THIS VERSION OF THE PROTOCOL.   
[electronic signature accepted]  
 
Principal Investigator:    
                     Date:  08/Feb/2023 
 
Statistical Co-investigator:   Agathe Ceppe  Date: 1/26/2021 
 
 

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Table of Contents 
Statement of Compliance .................................................................................................................................... 6 
Abbreviations and Definitions of Terms .............................................................................................................. 7 
1. 
Protocol Synopsis ..................................................................................................................................... 9 
2. 
Introduction: Background and Scientific Rationale .............................................................................. 11 
2.1. 
Background Information .................................................................................................................... 11 
2.2. 
Supporting Pilot / Unpublished Data ................................................................................................. 12 
2.3. 
Scientific Rationale ............................................................................................................................. 12 
3. 
Objectives .............................................................................................................................................. 12 
3.1. 
Specific Aim 1 ..................................................................................................................................... 12 
3.2. 
Specific Aim 2 ..................................................................................................................................... 12 
3.3. 
Specific Aim 3 ..................................................................................................................................... 12 
3.4. 
Specific Aim 4 ........................................................................................ Error! Bookmark not defined. 
4. 
Study Design .......................................................................................................................................... 13 
4.1. 
Treatment Design ............................................................................................................................... 13 
4.2. 
Experimental / Observational Design ................................................... Error! Bookmark not defined. 
4.3. 
Measurement Design ............................................................................ Error! Bookmark not defined. 
4.4. 
Outcome Measures ............................................................................................................................ 14 
5. 
Study Participants .................................................................................................................................. 14 
5.1. 
Number of Participants ...................................................................................................................... 14 
5.2. 
Eligibility ............................................................................................................................................. 14 
5.3. 
Strategies for Recruitment and Retention ......................................................................................... 16 
5.4. 
Consent Process ................................................................................................................................. 16 
6. 
Study Intervention ................................................................................................................................. 17 
6.1. 
Intervention – Test Article (if applicable) ........................................................................................... 17 
6.2. 
Assignment Procedures...................................................................................................................... 17 
6.3. 
Intervention – Procedural (if applicable) ........................................................................................... 17 
6.4. 
Concomitant Therapy (if applicable) .................................................................................................. 18 
6.5. 
Rescue Medications and Procedures (if applicable) .......................................................................... 18 
6.6. 
Compliance Checks ............................................................................................................................ 18 
6.7. 
Withdrawal / Discontinuation of Enrolled Participants ..................................................................... 18 
6.8. 
Voluntary Withdrawal (Drop-Out) of Enrolled Participants ............................................................... 18 
7. 
Study Procedures and Schedule ............................................................................................................ 19 
7.1. 
Table of Events ................................................................................................................................... 19 
7.2. 
Pre-Screening  / Screening  (Day -X to Day Y) .................................................................................... 19 

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7.3. 
Enrollment / Baseline (Day 0) ............................................................................................................ 19 
7.4. 
Study Visits (Numbered as V1, V2, etc., Day X ± Y) ............................................................................ 20 
7.5. 
Phone Contact (Day Day X ± Y) ........................................................................................................... 20 
7.6. 
Final Visit (Day Day X ± Y) ................................................................................................................... 20 
7.7. 
Follow-Up Contact (Day Day X ± Y) .................................................................................................... 21 
7.8. 
Premature Discontinuation ................................................................................................................ 21 
7.9. 
Collection and Management of Tissue Specimens ............................................................................ 21 
8. 
Study Measurements and Evaluations ................................................................................................. 21 
8.1. 
Outcome Measures for Evaluation of Feasibility / Tolerability .......................................................... 22 
8.2. 
Outcome Measures for Evaluation of Efficacy ................................................................................... 23 
8.3. 
Outcome Measures for Evaluation of Safety ..................................................................................... 23 
8.4. 
Other Outcomes in the Causal Pathway ............................................... Error! Bookmark not defined. 
8.5. 
Baseline Characteristics of the Participants .......................................... Error! Bookmark not defined. 
8.6. 
Variables Representing Treatment ....................................................... Error! Bookmark not defined. 
9. 
Statistical Analysis Plans........................................................................................................................ 23 
9.1. 
Strategies that Apply to all the Specific Aims..................................................................................... 23 
9.2. 
Description of the Study Cohort......................................................................................................... 23 
9.3. 
Aim-Specific Plans .............................................................................................................................. 24 
9.4. 
Planned Interim Analyses (If Applicable) ........................................................................................... 25 
9.5. 
Plans for Coping with Withdrawals and Loss-To-Follow-Up .............................................................. 25 
9.6. 
Sample Size Rationale ........................................................................................................................ 25 
10. 
Safety Monitoring and Management .................................................................................................... 26 
10.1. 
Risk / Benefit Assessment .................................................................................................................. 26 
10.2. 
Assessment of Safety ......................................................................................................................... 26 
10.3. 
Unanticipated Problems, Adverse Events, Serious Adverse Events ................................................... 27 
10.4. 
Safety Monitoring .............................................................................................................................. 28 
10.5. 
Study Suspension / Early Termination of the Study ........................................................................... 28 
11. 
Supporting Documentation and Operational Considerations ............................................................. 28 
11.1. 
Regulatory, Ethical, and Study Oversight Considerations .................................................................. 28 
11.1.1. 
Informed Consent Process ............................................................................................................. 29 
11.1.1.1. 
Consent/Assent and Other Informational Documents Provided to Participants ...................... 29 
11.1.1.2. 
Consent Procedures and Documentation .................................................................................. 29 
11.2. 
Study Discontinuation and Closure .................................................................................................... 30 
11.3. 
Confidentiality and Privacy ................................................................................................................ 31 
11.4. 
Future Use of Stored Specimens and Data ........................................................................................ 31 

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11.5. 
Key Roles and Study Governance ....................................................................................................... 32 
11.6. 
Safety Oversight ................................................................................................................................. 32 
11.7. 
Clinical Monitoring ............................................................................................................................. 33 
11.8. 
Quality Assurance and Quality Control .............................................................................................. 33 
11.9. 
Data Handling and Record Keeping .................................................................................................... 33 
11.9.1. 
Data Collection and Management Responsibilities ....................................................................... 33 
11.9.2. 
Study Records Retention ................................................................................................................ 33 
11.10. 
Protocol Deviations ........................................................................................................................ 34 
11.11. 
Publication and Data Sharing Policy  .............................................................................................. 34 
11.12. 
Conflict of Interest Policy ............................................................................................................... 34 
12. 
Additional Considerations ..................................................................................................................... 34 
13. 
References ............................................................................................................................................. 34 
14. 
List of Appendices .................................................................................................................................. 36 
 
 
 
 

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Statement of Compliance 
This study will be conducted in accordance with the International Conference on Harmonisation 
guidelines for Good Clinical Practice (ICH E6) and the Code of Federal Regulations on the Protection of 
Human Subjects (45 CFR Part 46). The statistical analysis plans will be consistent with guidances such as 
the CONSORT Statement [1] or STROBE Statement [2], ICMJE recommendations [3], the 2016 and 2019 
statements of the American Statistical Association [4,5], and recommendations in Nature [6,7].1 All 
personnel involved in the conduct of this study have completed human subjects protection training. 
 
 
 
 
1 [1] www.consort-statement.org [2] www.strobe-statement.org [3] www.icmje.org [4]  Wasserstein RL, et al. 
(2016), The ASA's Statement on p-Values, The American Statistician, 70:2, 129-133 [5]  Wasserstein RL, et al. 
(2019), Moving to a World Beyond p < 0.05, The American Statistician, 73:sup1, 1-19 [6]  Amrhein, et al. (2019) 
Scientists rise up against statistical significance, Nature 567, 305-307 [7] Editorial (2019) It’s time to talk about 
ditching statistical significance: Looking beyond a much used and abused measure would make science harder, but 
better. Nature 567, 283-283. 

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Abbreviations and Definitions of Terms 
 
Abbreviation/Acronym 
Definition 
AE 
Adverse Event/Adverse Experience 
AED 
Automated External Defibrillator 
ATS 
American Thoracic Society 
Ave60Clr 
Average Clearance at 60 minutes 
Ave120Clr 
Average Clearance at 120 minutes 
BP 
Blood Pressure 
CC 
Cough Clearance 
CEMALB 
Center for Environmental Medicine, Asthma and Lung Biology  
CF 
Cystic Fibrosis 
CI 
Confidence Interval 
CIOMS 
Council for International Organizations of Medical Sciences 
Co57 
Cobalt 57 
CONSORT 
Consolidated Standards of Reporting Trials 
C/P 
Central/Peripheral 
CRF 
Case Report Form 
CRO 
Contract Research Organization 
CTCAE 
Common Terminology Criteria for Adverse Events 
DSMB 
Data and Safety Monitoring Board 
EMS 
Emergency Medical Services 
eCRF 
Electronic Case Report Form 
FDA 
Food and Drug Administration 
FEF 25-75 
Maximal Mid-Expiratory Flow Rate 
FEV1 
Forced Expiratory Volume in one second 
FVC 
Forced Vital Capacity 
GCP 
Good Clinical Practice 
HIPAA 
Health Insurance Portability and Accountability Act 
ICMJE 
International Committee of Medical Journal Editors 
IRB 
Institutional Review Board 
IUD 
Intra Uterine Device 

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HR 
Heart Rate 
KeV 
Kilo electron volts 
LAIV 
Live Attenuated Influenza Vaccine 
N 
Number (typically refers to subjects) 
MCC 
Mucociliary Clearance 
mCi 
Millicurie 
MDI 
Metered Dose Inhaler 
MMAD 
Mass Median Aerodynamic Diameter 
NHLBI 
National Heart, Lung and Blood Institute 
NIH 
National Institute of Health 
PE 
Physical Exam 
PEF 
Peak Expiratory Flow 
PFT 
Pulmonary Function Test 
PHI 
Protected Health Information 
PI 
Principal Investigator 
RR 
Respiratory Rate 
QA 
Quality Assurance 
QC 
Quality Control 
SABA 
Short Acting Beta Agonist 
SAE 
Serious Adverse Event/Serious Adverse Experience 
SD 
Standard Deviation 
SE 
Standard Error 
SOP 
Standard Operating Procedures 
SpO2 
Peripheral capillary oxygen saturation 
STROBE 
STrengthening Reporting of OBservational studies in Epidemiology 
Tc99m-SC 
Technetium99m - Sulfur colloid 
uCi 
Microcurie 
UP 
Unanticipated Problem 
 

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1. Protocol Synopsis 
Study Title 
Functional Studies of Novel Genes Mutated in Primary Ciliary Dyskinesia II: 
Genotype to Phenotype 
Objectives 
Primary 
To measure mucociliary clearance (MCC) in subjects with primary ciliary 
dyskinesia (PCD) caused by mutations in genes associated with a milder clinical 
phenotype and subjects with PCD caused by mutations in genes associated with a 
more severe clinical phenotype. To compare the 2 groups with each other and 
with healthy individuals. 
Secondary 
To measure the effect of a short acting beta adrenergic (SABA) on MCC in in 
subjects with PCD caused by mutations in genes associated with a milder clinical 
phenotype and subjects with PCD caused by mutations in genes associated with a 
more severe clinical phenotype. 
Tertiary/Exploratory 
To measure cough clearance in subjects with PCD caused by mutations in genes 
associated with a milder clinical phenotype and subjects with PCD caused by 
mutations in genes associated with a more severe clinical phenotype. 
Target 
Population 
The target population is patients with PCD having confirmed mutations in genes 
associated with a milder clinical phenotype and subjects with PCD caused by 
mutations in genes associated with a more severe clinical phenotype. Subjects 
for this study will be recruited based on genotype. 
Key Inclusion Criteria  
       PCD Patients 
1. PCD subjects with mutations in genes associated with a milder clinical 
phenotype. This includes subjects with 2 confirmed mutations in RSPH1, 
RSPH9, RSPH4a, and DNAH11.This group may also include subjects with 
mutations in newly identified genes that are associated with a milder 
clinical phenotype. 
              OR 
PCD subjects with mutations associated with a more severe clinical 
phenotype. This includes subjects with 2 confirmed mutations in DNAH5, 
DNAI1, CCDC39, and CCDC40.This group may also include subjects with 
mutations in newly identified genes that are associated with a more 
severe clinical phenotype. 
2. Age 12 - 90 years old 
3. Subjects of childbearing potential must be non-pregnant and non-
lactating.  
Healthy Controls 
1. ≥ 18 years old 
2. No pre-existing lung disease (asthma, cystic fibrosis, etc.) 
4. Subjects of childbearing potential must be non-pregnant and non-
lactating.  

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Key Exclusion Criteria  
1. Clinically severe disease (requiring oxygen; FEV1<30%) 
2. Currently smoking or vaping 
3. Radiation exposure in the past year that would cause them to exceed 
federal radiation safety guidelines 
4. Free of sino-nasal pulmonary exacerbation for 4 weeks.  
Numbers of 
Participants 
Number to be recruited for screening : 40 
Number of eligible participants enrolled: 32 
Clinical Phase 
I 
Intervention 
Short acting beta agonist inhaler (albuterol by metered dose inhaler)  
Study 
Description 
Participants will inhale aerosolized radiolabeled sulfur colloid (Tc99m-SC) and will 
undergo gamma scintigraphy to measure mucociliary and cough clearance 
(MCC/CC) of the radiolabeled particles from the airways.  During the first hour of 
MCC imaging subjects will attempt to avoid coughing. Subjects will then inhale 4 
puffs of albuterol (90 mcg/puff via metered dose inhaler (MDI) with spacer) to 
assess the effect of beta-adrenergic stimulation on MCC over the second hour of 
MCC imaging. After the two hours of MCC imaging.  Subjects will perform 
spirometry and directed coughing over a 30-minute period during which CC will 
be measured, again by gamma scintigraphy. Subjects will then rest 
(approximately 3.5 hours) and be scanned for 15 minutes at 6 hour post initial 
MCC/deposition scan. 
Outcome 
Measures 
Baseline MCC: Ave60Clr 
Change in MCC following albuterol: Ave120Clr-Ave60Clr 
Change in MCC following cough clearance: Ave150Clr-Ave120Clr 
Study Duration 
The entire study is expected to last up to 4 years. 
Subject 
Participation 
Duration 
Each subject’s participation will last 1 day. 
Estimated Time 
to Complete 
Enrollment 
4 years  
Statistical 
Analysis Plans 
Aim 1 will provide estimates of the mean and standard deviation of 
baseline MCC (Ave60Clr) for each group of PCD subjects. The 2 disease 
groups will be compared to data from healthy subjects using ANOVA 
followed by multiple comparisons, using the Tukey Kramer correction. Age, 
gender and the initial deposition C/P will be added as covariates to PCD 
groups in a linear regression model on MCC. 
 
In Aim 2, the change pre- to post-albuterol (Ave120Clr-Ave60Clr) will be 
calculated for each subject. Estimates of the mean and standard 
deviations will be reported for each group and compared with paired t-
tests. The difference of change between the 2 groups will first be analyzed 
with an independent t-test. Age, gender and C/P will then be added as 
covariates in linear regression model. 

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In Aim 3, the change pre- to post-cough clearance (Ave150Clr-Ave120Clr) 
will be calculated for each subject. Estimates of the mean and standard 
deviations will be reported for each group and compared with paired t-
tests. The difference of change between the 2 groups will be analyzed with 
an independent t-test. Age, gender and C/P will then be added as 
covariates in linear regression model. 
 
All tests will be 2-sided, with α=0.05. Tests with p values above 0.05 will 
be considered inconclusive. 
2. Introduction: Background and Scientific Rationale 
2.1. Background Information 
Primary Ciliary Dyskinesia (PCD) is a heterogeneous genetic disease that results in impaired function of 
the cilia [1]. Pulmonary MCC is dependent on airway secretory cells and submucosal glands and ciliated 
cells to propel mucus upwards through the airways. Rates of MCC are dependent on ciliary beat 
frequency, hydration, and the rheologic properties of mucus [2]. Abnormalities in ciliary function causes 
reduction in mucociliary clearance (MCC), which contributes to mucus plugging and worsening of lung 
function. Due to cilia dysfunction, mucociliary clearance (MCC) and cough clearance (CC) are affected in 
PCD. PCD varies in phenotypic presentation and ranges from mild to severe loss of lung function. PCD 
patients develop airway obstruction and bronchiectasis as disease advances. While some patients 
present with bronchiectasis in childhood, virtually all adult patients have bronchiectasis [3]. PCD is a 
progressive disease causing a range of respiratory disease from poor lung function to chronic respiratory 
failure requiring lung transplant [3]. 
As more genetic mutations are discovered, milder phenotypes have been associated mutations in 
certain genes. For example, it has been reported that individuals with mutations in RSPH1 (radial spoke 
head homolog 1) have a milder presentation than other genes [4]. Individuals with RSPH1 have a higher 
nasal nitric oxide value, decreased incidence of neonatal respiratory distress and higher FEV1 when 
compared to subjects with mutations in genes encoding outer dynein arm structures. In addition, cilia 
from subjects with mutations in RSPH1 can have normal appearing structure and maintain an almost 
normal ciliary beat frequency, although they often beat in a circular pattern [5]. In contrast, mutation in 
other genes have been associated with a more severe clinical phenotype. For example, subjects with 
mutations in CCDC39 or CCD40 were significantly younger at diagnosis and had significantly lower 
percent predicted FEV1 than subjects with mutations in other genes [6]. 
Currently, there are no treatments that have been clinically approved specifically for PCD. In other 
disease processes, albuterol has been reported to improve mucociliary clearance. Laube et al reported 
decreased MCC in patients after lung transplant which significantly improved with administration of 
albuterol [7]. Additionally, beta agonists have been reported to improve ciliary beat frequency in human 
bronchial epithelial cells in vitro and in certain animal models [8-9]. The effect of beta agonists on MCC 
or CC has not been studied in those with mild or severe phenotypes of primary ciliary dyskinesia. A 
recent retrospective study did investigate bronchodilator response (BDR) with a SABA in patients with 

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PCD (adults and children), and reported a higher prevalence of BDR in adults and those with defects of 
the microtubule compared to younger patients and those with other defects [12]. 
 
2.2. Supporting Pilot / Unpublished Data 
N/A 
 
2.3. Scientific Rationale 
We have previously reported that PCD patients with RSPH1 mutations have a milder pulmonary 
phenotype and better lung function than patients with PCD caused by other mutations [3]. Further, we 
demonstrated that although cilia lacking RSPH1 beat in an abnormal, circular pattern, Rsph1-/- mice 
exhibited a low level of MCC [13]. This is in contrast to Dnaic1-/- mice, who exhibit essentially immotile 
cilia and a complete lack of MCC [14]. In this aim, we will test the hypothesis that PCD subjects with 
mutations in RSPH1 and certain other mutations that have a milder phenotype [ 4, 11] maintain a low 
level of MCC that is likely responsible for their milder disease phenotype. In addition, we will test the 
hypothesis that stimulating ciliary beat frequency (CBF) with a beta-agonist will increase MCC in this 
patient group. These studies may result in the development of new personalized treatments for this rare 
disease. 
3. Objectives 
3.1. Specific Aim 1 
To estimate how MCC measurements vary in subjects with PCD caused by mutations in genes associated 
with mild and severe clinical phenotypes relative to healthy subjects, and compare the 3 populations in 
terms of Average Clearance.  
3.2. Specific Aim 2 
To characterize the effect of a short acting beta agonist (SABA) on MCC subjects with PCD caused by 
mutations in genes associated with mild and severe clinical phenotypes and compare the 2 populations 
in terms of change in Average Clearance. 
3.3. Specific Aim 3 
To characterize effect of cough clearance in subjects with PCD caused by mutations in genes associated 
with mild and severe clinical phenotypes and compare the 2 populations in term of change in Average 
Cough Clearance. 
 
 
 

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4. Study Design 
 
 
Prior to  
Enrollment 
 
                                                        
                                                              
                                       
 
 
 
 
 
 
 
 
 
 
 
4.1 Treatment Design 
Type of Design: Open label; two group study with healthy controls 
Screening/Baseline: Participants will undergo a general medical screening visit to include vital signs, 
physical exam and pregnancy test (if applicable), then proceed with spirometry. 
Intervention/Treatment 
Participants will inhale aerosolized radiolabeled sulfur colloid (Tc99m-SC) and will undergo gamma 
scintigraphy to measure mucociliary and cough clearance (MCC/CC) of the radiolabeled particles from 
the airways in both groups.  During the first hour of MCC imaging patients will attempt to avoid 
coughing. Patients will then inhale 4 puffs of albuterol (90 mcg/puff via metered dose inhaler (MDI) with 
spacer) to assess the effect of beta-adrenergic stimulation on MCC over the second hour of MCC 
Total N:  Obtain informed consent over the phone or by virtual meeting. Screen 
potential participants by inclusion and exclusion criteria; obtain medical history, 
document over the phone or by virtual meeting. 
Perform screening and spirometry. Perform mucociliary (MCC) baseline 
measurement. Continue MCC measurement after SABA administration. Repeat 
spirometry followed by controlled cough maneuvers to assess cough clearance 
(CC). Subject will rest for approximately 3.5 hours followed by a final MCC/CC static 
measurement, 6 hours (±30 minutes) post initial scan.  
Arm 1  
Mild Mutation 
N =12 
Arm 2 
Severe Mutation 
N =12 
Arm 3 
Healthy Controls 
N=8 

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imaging. After the two hours of MCC imaging patients will perform spirometry and directed coughing 
over a 30 minute period during which CC will be measured, again by gamma scintigraphy. Subjects will 
then rest (app 3.5 hours) and be scanned for 15 minutes at 6 hour ±30 minutes post initial 
MCC/deposition scan. 
Table of procedures 
 
Unscheduled Visits: There is no expectation for unplanned visits.  Subjects will simply be rescheduled if 
they have a change in health.   
 
4.1. Outcome Measures 
Primary Outcome(s):  Baseline MCC (Ave60Clr) in subjects with PCD caused by mutations associated 
with mild and severe clinical phenotypes. 
Secondary Outcome(s): Change in MCC (Ave120Clr-Ave60Clr) after the administration of albuterol in 
subjects with PCD caused by mutations associated with mild and severe clinical phenotypes PCD 
patients with mild disease compared to PCD patients with more severe disease. 
Tertiary/Exploratory Outcome(s): Change in cough clearance (Ave150Clr-Ave120Clr) in subjects with 
PCD caused by mutations associated with mild and severe clinical phenotypes PCD patients with mild 
disease compared to PCD patients with more severe disease. 
5. Study Participants 
5.1. Number of Participants 
Number to be recruited for screening: 40 
Number of eligible participants enrolled  32 
5.2. Eligibility 
 
Inclusion Criteria for PCD Patients 
1. Age 12-90 of both genders 

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2. Confirmed PCD diagnosis with identified genetic mutations.  
3. Negative pregnancy test for females who are not s/p hysterectomy with oophorectomy. 
4. FEV1 of at least 30% of predicted. 
 
Inclusion Criteria for Healthy Controls: 
1. Age ≥ 18 years old. 
2. Subjects must have an FVC, FEV1 and FVC/FEV1 of at least 80% of predicted.  Subjects who fall 
out of the normal range will be offered a copy of the test to share with their personal physician. 
3. No pre-existing lung disease (asthma, cystic fibrosis, etc.). 
4. Negative pregnancy test for females who are not s/p hysterectomy with oophorectomy. 
 
Exclusion Criteria for all participants:  
Clinical Contra-indications:  
1.  Any chronic medical condition considered by the PI as a contraindication to the exposure study 
including significant cardiovascular disease, diabetes, chronic renal disease, chronic thyroid disease, 
immunodeficiency, history of tuberculosis  
2. Any acute infection requiring antibiotics within 4 weeks of study.  
3. Mental illness or history of drug or alcohol abuse that, in the opinion of the investigator, would 
interfere with the participant’s ability to comply with study  
requirements. 
4. Medications which may impact the results of the study treatment, or may  
interfere with any other medications potentially used in the study (to include  
steroids, beta antagonists, non-steroidal anti-inflammatory agents)  
5. Active smoking to include e-cigarettes within 1 year of the study, or lifetime of  
> 10 pack years of smoking 
6. History of vaping or current vaping.  
7. Allergy/sensitivity to study drugs, or their formulations.  
8. Viral upper respiratory tract infection within 4 weeks of challenge.  
9. Radiation exposure history in the past year which would be outside the safe  
levels  
10. Pregnant or lactating women will also be excluded since the risks associated with radiation are 
unknown and cannot be justified  
11. Use of the following medications:  
a. Use of beta blocking medications 
b. Receipt of LAIV (Live Attenuated Influenza Vaccine), also known as FluMist , within the prior 
30 days, or any vaccine within the prior 5 days  
c. Multivitamins, Vitamin C or E or herbal medications in the 4 days prior to the treatment visit 
d. Non-steroidal anti-inflammatory drugs in the 4 days prior to the treatment visit. 
12. Known IgE mediated hypersensitivity to albuterol 

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13. Physical/laboratory indications:  
a. Temperature > 37.8  
b. Subjects >15 y.o - Systolic BP >150 mm hg or < 90 mm Hg or diastolic BP> 90 mm Hg or < 50 
and Subjects 12-15 y.o – Systolic BP > 130 mmHg or < 80 mmHg or diastolic BP > 80 or <40 
c. Oxygen saturation of < 93%  
14. Inability or unwillingness of a participant to give written informed consent. 
 
5.3. Strategies for Recruitment and Retention 
Recruitment Strategy:  
We will recruit subjects who have a known diagnosis of PCD caused by mutations in genes associated 
with a milder clinical phenotype (RSPH1, RSPH9, RSPH4a, and DNAH11) and subjects with PCD caused by 
mutations in genes associated with a more severe clinical phenotype (DNAH5, DNAI1, CCDC39, and 
CCDC40). In addition, we will recruit healthy controls from the general public to include fliers around 
UNC campus, e-mails, and/or by word of mouth.   PCD subjects may be recruited from pediatric or adult 
PCD clinics at UNC by physicians listed on the protocol including Dr. Peader Noone, Dr. Margaret Leigh, 
and Dr. Sara Abu-Nasser. In addition, subjects may have had diagnostic information obtained from other 
research studies completed at our institution and will be recruited from previous PCD research studies. 
The research studies below will provide diagnostic PCD information for patients: 98-1015 (1395), 05-
2997 (5901), 05-2979 (5902), 08-0764 (5903), 10-1523 (5904), 14-1225 (5905), and 20-0508 (5906). 
Once patients with the above genetic mutations are identified, patients will be called by a study 
coordinator or a study physician to recruit subjects for the study.  
Retention Strategy:  
We will perform the initial screening over the phone or virtually, in order to decrease the number of in 
person visits required. The study is to be performed over the span of one day, and will not require 
follow-up, therefore it is less likely for subject to drop out.  
Screen Failures:  
For subjects who do not meet enrollment criteria, all study related activity will stop once it is 
determined that the subject does not meet criteria. 
5.4. Consent Process 
Once subjects have been identified, a study coordinator or physician will call the patient to present and 
describe the study. Due to COVID-19 pandemic, precautions to limit in person contact include using 
phone calls or virtual calls to explain and discuss the study. The study coordinator/physician will also 
review the consent form with the subjects in the same method if they agree to the study. If subject 
agrees, a signature for the consent form will be required on the day of the study.   
For children under the age of 18, their guardian or parent will be approached in the same manner as 
above. After explanation of the study, the consent form will be explained to the parent or guardian. 
Parent or guardian will sign the consent form the day of the study. The study procedures will also be 

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explained to the subject in age appropriate language during the call.  Written and signed assent will be 
obtained the day of the procedure. When reasonably available, two parent or guardians will be required 
to provide consent for minors to participate.  For situations when a parent is available able to consent 
but obtaining the written signature is not feasible, DocuSign may be used to document parental 
permission electronically. 
 
6. Study Intervention 
6.1. Intervention – Test Article  
Description: Albuterol; HFA 90 mcg/puff; 4 puffs; one time. It is available commercially as ProAir HFA or 
Ventolin HFA. 
Formulation/Packaging/Labeling: [As supplied by manufacturer/Sponsor] 
Open label study 
Storage and Stability:  
Albuterol MDI will be delivered to and stored at a secure location within the research office. Albuterol 
MDI is stable at room temperature.  
Preparation and Administration:  
Albuterol MDI inhaler will require a spacer to be administered. Dose is 4 puffs one time for patients of 
all ages.  
Modification:  
No anticipated modification necessary. 
Accountability:  
Albuterol MDI inhaler will be disposed of after the completion of the study.  
6.2. Assignment Procedures 
Matching and Stratification: N/A 
Randomization, Concealment, and Blinding: (N/A) 
Masking (Blinding): N/A 
 
6.3. Intervention – Procedural (if applicable) 
Description: [N/A 
Training on Procedural Intervention: N/A 

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6.4. Concomitant Therapy (if applicable) 
N/A 
6.5. Rescue Medications and Procedures (if applicable) 
N/A 
6.6. Compliance Checks 
N/A 
6.7. Withdrawal / Discontinuation of Enrolled Participants 
Patient’s participation will be discontinued if: 
1. Subject develops new or worsening respiratory symptoms including shortness of 
breath or decrease in FEV1 of 20% or greater.  
2. Subject develops a condition that is an exclusion criterion. 
3. Subject is noncompliant with the recommended withholding of SABA or ICS/LABA 
for 12 hours prior to study. 
If any of the above are met, then patient will not continue. 
 
6.8. Voluntary Withdrawal (Drop-Out) of Enrolled Participants 
Participants may voluntarily withdraw participation at any time, for any reason, with no penalty or loss 
of rights. Patients will not require follow up if they withdraw and can continue routine medical care. 
 
 
 
 
 
 
 
 
 
 
  

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7. Study Procedures and Schedule 
7.1. Table of Events 
Procedures 
 
MCC Visit 
(Day 1) 
Informed Consent 
 
X 
Medical History, 
med review 
 
X 
Concomitant 
Medications  
 
X 
COVID-19 Screen 
 
X 
Physical Exam 
Complete 
X 
Symptom-Directed 
X 
Vital Signs 
X 
Urine Pregnancy 
Test 
 
X 
Spirometry 
 
X 
MCC 
 
X 
 
 
7.2. Pre-Screening  / Screening   
Subjects will be screened by phone for potential eligibility and interest once they are identified using the 
recruitment strategies discussed in Section 5.3. Screening will review inclusion and exclusion criteria.  
Approximately 48 hours before the study visit, subject will be screened for COVID-19 using the screening 
questions stated in section 7.4 and reminded to withhold use of SABA or LABA 12 hours prior to visit, and 
airway clearance medications (e.g. Hypetonic Saline or Pulmozyme) for 24 hours before the visit.  
7.3. Enrollment / Baseline (Day 0) 
On the day of the study, medical history and exclusion criteria will be reviewed again. Consent will also 
be signed on day of the study.  
 
 
 
 

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7.4. Study Visits  
Intervention/Treatment procedures for MCC (by visits)  
Adverse events are assessed at each visit.  
Screening Call: Approximately 48 hours before the study, patient will undergo a COVID-19 screen with 
the questions below: 
Do you feel feverish or have a fever? 
Do you have a new or worsening cough? 
Do you have a new or worsening sore throat? 
Are you newly short of breath? 
Have you had a new onset of loss of taste or smell in the last 5 days? 
Do you have new onset vomiting or diarrhea? 
Do you have new onset of repeated shaking with chills not related to another medical condition?  
If patient answer yes to any of the questions above, study will be cancelled. 
 
MCC/Treatment visit: On day of the study, medical history will be updated, including any new 
medications or new medical history. Vital signs and baseline spirometry will be collected, and a negative 
urine pregnancy test will be required for all women.   
Participants will inhale aerosolized radiolabeled sulfur colloid (Tc99m-SC) and will undergo gamma 
scintigraphy to measure mucociliary and cough clearance (MCC/CC) of the radiolabeled particles from 
the airways.  During the first hour of MCC imaging patients will attempt to avoid coughing. Subjects will 
then inhale 4 puffs of albuterol (90 mcg/puff via metered dose inhaler (MDI) with spacer) to assess the 
effect of beta-adrenergic stimulation on MCC over the second hour of MCC imaging. After the two hours 
of MCC imaging, patients will perform spirometry and directed coughing over a 30-minute period during 
which cough clearance will be measured, again by gamma scintigraphy. Subjects will then rest (app 3.5 
hours) and be scanned for 15 minutes at 6 hour (±30 minutes) post initial MCC/deposition scan. 
All personnel performing aerosolizing procedures (MCC scans and spirometry) will wear eye protection 
and N95 masks. A HEPA filter will also be placed in the room where the mucociliary scans will be 
performed. 
 
7.5. Phone Contact (Day Day X ± Y) 
N/A 
 
7.6. Final Visit (Day Day X ± Y) 
N/A 
 

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7.7. Follow-Up Contact (Day Day 1 ± 3) 
The study team will contact participants between 1 and 4 days after the study visit to confirm when 
study payment card has been loaded and to follow up on any questions or concerns. 
7.8. Premature Discontinuation 
Study is considered completed after the last scan of the day. Subjects may withdraw at any time. 
Subjects may be withdrawn at any time for safety concerns. Subjects who have any unanticipated 
problems (UPs) with the study procedures will be monitored until resolution of the symptoms. 
 
7.9. Collection and Management of Tissue Specimens 
Sample Preparation: N/A 
Record Keeping and Monitoring: N/A 
Sample Storage and Security: N/A 
8. Study Measurements and Evaluations 
 
List variables that will be abstracted from medical charts: Medical records will be  
reviewed to obtain previous FEV1 measurement and to confirm concomitant medications.  
Describe screening evaluation: Vital signs, including heart rate, respiratory rate, temperature and blood 
pressure will be collected. Oxygen saturation levels will be noted, as well as breath sounds. A brief physical 
examination, including but not limited to the cervical lymph nodes, eyes, ears, nose, throat, cardiovascular 
and respiratory systems will be performed.  
 
Describe how measurements will be taken: 
Spirometry:  
This test measures the volume of air that can be exhaled and the rate of airflow during exhalation after a 
maximal inhalation and is performed to American Thoracic Society (ATS) guidelines. Subjects will inhale as 
deeply as possible, then exhale as rapidly and completely as possible into the spirometer. Measurements 
obtained from each maneuver include the forced vital capacity (FVC), the forced expiratory volume in the 
first second (FEV1), the maximal mid-expiratory flow rate (FEF 25-75%) and the peak expiratory flow (PEF). 
The largest FVC and FEV1, from at least 3 acceptable trials, are selected for analysis; the flow rates are 
selected from the trial with the largest sum of FVC and FEV1.  
 
Albuterol Inhalation:  
The subject will be instructed on the proper use of MDI Inhaler, an approved device for inhalation of 
albuterol. The subject will use the inhaler to administer 4 puffs of albuterol.  
 

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MCC procedures:  
Prior to each MCC study, a transmission Co57 scan will be performed to define the lung boundaries, to assign 
regions of interest, and to normalize these regions for lung volume differences. A rectangular phantom 
containing the radioisotope Co57 (< 25 mCi) will be placed in front (5cm) of the subject sitting with his/her 
back to the gamma camera for 30 seconds. The transmission scan has been used by us (e.g., 05-2358, 08-
0795, 06-1016, 13-1605, 15-938) and others to provide a delineation of lung boundaries for generating the 
central (C) and peripheral (P) regions of interest (ROIs) used to assess initial particle deposition (C/P ratio) 
and subsequent clearance of the inhaled radio-aerosol [15-16]. Prior to the transmission scan on each study 
day, we will place 2 spot markers of Americium241 (0.9 microcurie (uCi) each, gamma 66 KeV) on the upper 
and lower back of each subject during scanning (both Tc99m-SC deposition/retention and Co57 
transmission). With dual isotope imaging, these spot markers will allow alignment of images for more 
accurate determination of regional deposition/retention. These very low radiation sources have been 
obtained from commercially available home smoke alarms. The placement of these markers will be 
determined to be outside the lung field during the transmission scan. Their location will be marked in semi-
permanent ink for later placement during Tc99m deposition/retention scans. The shielded side of this source 
will be placed/taped onto the subject’s skin. Radiolabeled Tc99m-sulfur colloid will be delivered using a 
modified Pari-LC Star nebulizer (MMAD 9.5 um). This is a closed delivery system that produces 80 ml/sec air 
flow, and therefore limits the inspiratory flow rate to this value. While seated in front of a gamma camera 
subjects will perform single inhalations lasting ~10 seconds each from the delivery system and will exhale at 
500 ml/sec (using feedback from a flow meter in the breathing circuit). Approximately 5 of these inhalation 
maneuvers will be required to deposit an adequate isotope dose to the lung. Subjects will be allowed to 
breathe normally (off the nebulizer) in between each inspiratory maneuver. Each volunteer will practice 
these maneuvers prior to the actual radio-aerosol inhalation to guarantee his/her proficiency. The activity of 
Tc99m-SC loaded in the nebulizer will be adjusted to provide an estimated 40 uCi deposited in the lung for 
the MCC/CC scan. Patients between the age 12-18 years old will receive ¾ of the adult dose to account for 
the smaller lung volume.  A single crystal detector will be placed at the subject’s back during inhalation to 
monitor dose to the lung. Total inhalation time should be less than 5 minutes in all cases. Immediately 
following isotope inhalation, the subject will gargle and drink water to clear activity that deposited in the 
mouth into the stomach. The subject will then (within a minute of final inhalation maneuver) be seated in 
front of a large-field-of-view gamma camera to begin acquiring consecutive 2 minute images. The first two-2-
minute images will provide initial, time zero activity (i.e. 100% retention) followed by the same imaging at the 
start of every 10-minute period until 1 hour has passed to assess baseline MCC. Subjects will then inhale 4 
puffs of albuterol from the MDI inhaler and consecutive 2 min imaging continues for the next hour to assess 
the effect of albuterol on MCC. PFTs will then be performed and subjects will cough a total of 30 times over a 
30-minute period to assess CC by gamma imaging over that period. Finally, a static 15-minute image will be 
measured at 6 hours (±30 minutes) after the initial radio- aerosol deposition scan. (See table of procedures in 
section 4.1). 
 
8.1. Outcome Measures for Evaluation of Feasibility / Tolerability 
N/A 
 

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8.2. Outcome Measures for Evaluation of Efficacy 
First estimates of the efficacy of SABA for accelerating MCC immediately in PCD subjects, will be 
obtained by comparing the baseline rate of MCC and the rate of MCC immediately after albuterol 
inhalation.  
 
8.3. Outcome Measures for Evaluation of Safety 
Vital signs are monitored throughout the study as needed, including heart rate, respiratory rate, 
temperature, and blood pressure.  
We will assess vital signs at baseline, and then as needed when indicated by study physician. 
 
9. Statistical Analysis Plans 
 
9.1. Strategies that Apply to all the Specific Aims 
To describe each measure of MCC by retention vs. time data set, the average % clearance (or 100* (1- 
Retention)) over the 0-60 and 60-120 min period of observation are computed (i.e. average of the 10 
minute clearance values from 0 to 60 and 60-120 minutes). These computed values are signified as 
Ave60Clr and Ave120Clr respectively. Each calculated AveClr value utilizes all 10-minute data points, 
rather than simply reporting a clearance value at a single time, and represents the average clearance at 
the midpoint of each period of retention vs. time observation. 6 hour clearance is determined by (100* 
(1- 6 hour retention)) and provides an index of initial airway (6hr Clr) vs. alveolar (6hr retention) deposition 
that along with the central/peripheral (C/P) ratio provides regional deposition data that can be included 
as covariates in analysis of MCC comparisons between groups and within individuals. 
Distributions of the outcomes is expected to be normal. We will plot the data in histograms and qqplot to 
evaluate any extreme datapoints or outliers. Their impact if any will be analyzed by performing the 
analysis with and without these datapoints. 
 
9.2. Description of the Study Cohort 
 
The study cohort will consist of 12 PCD subjects with mutations in genes associated with a mild 
pulmonary phenotype (RSPH1, RSPH9, RSPH4a, or DNAH11) and 12 PCD subjects with mutations in 
genes associated with a more severe pulmonary phenotype (DNAH5, DNAI1, CCDC39, or CCDC40). A 
subject with mild pulmonary phenotype will typically have later onset of respiratory symptoms, better 
lung function, and milder bronchiectasis [4].  The age of patients will range between 12-90 years old. 
Eight healthy patients will be enrolled and undergo the same MCC protocol to obtain data for 
comparison. All subjects following the protocol will be included in the database. Subjects not 

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completing the protocol will be replaced.  Baseline measurements of population (age, gender, race, 
FEV1) will be tabulated, for the whole population and per group (mild genotype, severe genotype, 
healthy) using mean and standard deviation and frequencies with percentages as appropriate. Data will 
be plotted in graphical figures including as histograms, bar plots, and scatter plots.  
 
9.3. Aim-Specific Plans 
Aim 1: To estimate how MCC measurements vary in PCD subjects with mutations in genes that are 
associated with mild or severe disease, relative to healthy subjects, and compare the 3 populations in 
term of Average Clearance. 
Aim 1 is measuring MCC in PCD subjects having confirmed mutations in genes associated with a milder 
clinical phenotype and subjects with PCD caused by mutations in genes associated with a more severe 
clinical phenotype. These measurements will be reported as estimates of the means and standard 
deviations of Ave60Clr for each group, with associated confidence intervals. The baseline MCC in PCD 
subjects with “mild genotypes” and “severe genotypes” will be compared to healthy subjects.  Our 
expected result is that subjects with mutations in genes associated with mild disease will display lower 
MCC rates than healthy subjects, and subjects with mutations in genes associated with severe disease will 
be even lower. For this comparison, we will use an ANOVA followed by multiple comparisons, using the 
Tukey Kramer correction. Then, to control for other possible sources of variations, age, gender and the 
initial deposition C/P will be added as covariates to PCD groups in a linear regression model on MCC. 
Aim 2: To characterize the effect of a short acting beta agonist (SABA) on MCC in PCD subjects with 
mutations in genes that are associated with mild or severe disease, and compare the 2 populations in 
terms of change in Average Clearance. 
In aim 2, we will be measuring the effect of albuterol on MCC in PCD subjects. We will look at the change 
between Ave60Clear (before albuterol) and Ave120Clr (after albuterol) for each subject. Estimates of the 
means and standard deviations with associated confidence intervals will be reported for each group. Using 
paired t-tests, we will verify if the change is significantly different than 0. The difference between the 2 
PCD groups will then be analyzed with an independent t-test. Age, gender and the initial deposition C/P 
will be added as covariates to PCD groups in a linear regression model on MCC. 
As this is not a randomized crossover design nor a parallel-arm design, our proposed comparison pre- to 
post-albuterol is uncontrolled and cannot assess the efficacy of SABA. The goal of our study is to obtain 
first estimates of the immediate effect albuterol on MCC in a small, controlled population.   
Aim 3: To characterize cough clearance in PCD subjects with mutations in genes that are associated with 
mild or severe disease, and compare the 2 populations in terms of change in Average Clearance. 
We will compare cough clearance in PCD subjects with mutations associated with mild disease with PCD 
subjects with mutations in genes associated with more severe disease. Cough clearance will be 
measured as the change in retention between 120 and 150 min post deposition. This change between 
Ave120Clr and Ave150Clr will be tabulated for each PCD group in term of means and standard 
deviations with associated confidence intervals and compared to 0 using paired t-tests. The difference 
between the 2 PCD groups will then be analyzed with an independent t-test. Age, gender and the initial 
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For all aims, all tests will be 2-sided, with α=0.05. Tests with p values above 0.05 will be considered 
inconclusive.  
Expected results, potential problems, and alternatives.  We expect to observe a significant rate of MCC 
(different from 0) in the group of PCD patients with mutations in genes associated with a milder 
phenotype. Based on previous studies of ciliary activity in PCD subjects with mutations associated with a 
more severe phenotype, we expect to see little or no MCC in this group of PCD patients. Further, we 
expect the rate of MCC to be significantly greater in subjects in the “mild genotype” group compared to 
the PCD patients in the “severe genotype” group. We expect that subjects with a mild genotype will 
respond to albuterol with an increase in MCC while the PCD patients with the severe genotype will not. 
We expect that subjects with mutations in genes associated with a mild phenotype will have better 
cough clearance compared to subjects with mutations in genes associated with a severe phenotype. As 
we have performed many similar studies in the past, including studies of MCC in PCD subjects, we 
anticipate no technical difficulties in carrying out these studies. If there is no difference detected 
between the groups of PCD patients, this would suggest that the less severe clinical phenotype observed 
in some patients is not due to the maintenance of a low level of MCC. Nevertheless, this would be an 
important experimental finding, as it would indicate that some other mechanism is responsible for the 
difference in phenotype. 
9.4. Planned Interim Analyses (If Applicable) 
N/A 
 
9.5. Plans for Coping with Screen Failures, Withdrawals and Loss-To-Follow-Up 
Participants will be screened for key eligibility criteria over the phone prior to the study visit. Only subjects 
that meet the key eligibility will be scheduled for a visit.  If any eligibility criteria is not met on day of the visit 
(eg. Vital signs, sprirometry results), subject will be considered a screen-failure and no additional procedures 
will be completed.  Additional participants will be recruited to replace screen failures in order to meet target 
enrollment numbers. 
Due to the short duration of the study, we do not except any withdrawal or loss to follow up patient. In 
the case a subject would withdraw consent, we will recruit another subject to reach our desired sample 
size for analysis.  
 
 
9.6. Sample Size Rationale 
Sample size:  
Based on the results of mucociliary transport (MCT) rates (healthy: 6.58 ± 1.39, Rsph1-/-: 1.63 ± 0.78, other 
PCD: 0) in our mouse model [13], we expect subjects with a mild genotype to average 25% of the MCC of 
healthy subjects (healthy: Ave60Clr =14.3 ± 7.2% ; mild expected: 3.6 ± 2%), while subjects with a severe 
genotype will not show any MCC results, i.e. Ave60Clr not different from 0 (expected 0.5 +/- 0.25). With 
n=10, we will be able to observe significant differences between all groups, with α=0.05 and power=0.8.  

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Moreover, we expect the mild genotype group to react to albuterol treatment by doubling their Ave60 
Clr, while the group with severe mutations will not. With 10 subjects, we will be able to find a significant 
response in the mild genotype group with expected Ave60Clr = 7.2 ± 4%. Accounting for 20% possible 
dropout, we plan to enroll 12 subjects. 
 
10. Safety Monitoring and Management 
 
10.1. 
Risk / Benefit Assessment 
Potential Risks:  
 
 
Potential Risks of Spirometry:  
Potential risks include possible lightheadedness or wheezing. 
 
Potential Risks of Mucociliary clearance scan:  
The radiation risks for the MCC scan, including the Cobalt 57 transmission scan and the 
Americium241 disks, which are used as fiducial markers is approximately 44 mRems. Adults in 
the New York City area receive about 300mRems per year in natural radiation exposure.  
 
Potential Benefits: 
 
Potential Benefits includes looking at possible treatment to improve mucociliary clearance and/or 
cough clearance in patients with PCD.  
 
10.2. 
Assessment of Safety 
 
The study procedures are low risk and are not expected to cause any adverse events, however, to ensure that 
subjects are tolerating the study well we have listed the screening and assessments below. General screen 
including physical exam at the start of the study and symptom-based exams during the study. 
• 
Spirometry will be completed at baseline and after administration of SABA.  
• 
Pulse oximetry will be checked at the start of the study and as needed. 
• 
Vital Signs will be obtained at the start of the study and as needed. 
 
 
Protections to Minimize Risk of Spirometry:  
- Subjects will be seated in a non-rolling chair when spirometry is performed and standard 
methodology conforming to the American Thoracic Society guidelines for measurement of 
spirometry will be used. Subjects will be instructed to notify the study staff if they feel lightheaded, 
and albuterol will be available in the event the subject experiences any unanticipated 

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bronchoconstriction.  
 
Protections to Minimize Risk of Mucociliary clearance scan:  
- Radiation history is collected. Any subject who will exceed Federal guidelines for safe annual 
exposure limits will not be enrolled. 
 
10.3. 
Unanticipated Problems, Adverse Events, Serious Adverse Events 
Definition of Adverse Event (AE) and Serious Adverse Event (SAE) 
An adverse event for a given volunteer will be defined as failure of any of the safety criteria outlined 
above. Decrease in lung function (FEV1) greater than 20% will be considered an adverse event. Any 
symptoms that induce a volunteer to seek medical attention from any provider within 96 hours of a study 
visit will be considered an adverse event. A serious adverse event will be defined as any event that 
requires hospitalization or results in life threatening illness or injury, permanent (or likely to be permanent) 
illness or injury, or death if these events occur within 96 hours of a study visit (or if the clinical scenario 
leading up to hospitalization, illness, injury or death begins within 96 hours of a study treatment visit).  
Grading criteria  
In addition to determining whether an adverse event fulfills criteria for a serious adverse event or not, the 
severity of adverse events experienced by study participants will be graded according to the criteria set 
forth in the National Cancer Institute’s Common Terminology Criteria for Adverse Events Version 4.0. This 
document (referred to herein as the NCI-CTCAE manual) provides a common language to describe levels 
of severity, to analyze and interpret data, and to articulate the clinical significance of all adverse events.  
 
All adverse events whether or not listed in the NCI-CTCAE will be graded on a scale from 1 to 5 
according to the following standards in the NCI-CTCAE manual (A semi-colon indicates ‘or’ within the 
description of the grade.):  
Grade 1 = Mild; asymptomatic or mild symptoms; clinical or diagnostic observations only; intervention not 
indicated. 
Grade 2 = Moderate; minimal, local or noninvasive intervention indicated; limiting age- appropriate 
instrumental ADL (preparing meals, shopping for groceries or clothes, using the telephone, managing 
money, etc). 
Grade 3 = Severe or medically significant but not immediately life-threatening; hospitalization or 
prolongation of hospitalization indicated; disabling; limiting self care ADL (bathing, dressing and 
undressing, feeding self, using he toilet, taking medications, and not bedridden). 
Grade 4 = Life-threatening consequences; or urgent intervention indicated. 
Grade 5 = Death related to AE.  
 
Adverse Event/Serious Adverse Event reporting procedures: Any SAE’s and UP’s will be reported to 
the IRB and the Medical Monitor. The funding agency will be notified of any SAEs.  
Medical Emergency procedures: There is a study physician available for all inhalation procedures. A 
code cart and Automated External Defibrillator (AED) are available in the event of a cardiopulmonary 

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event. Albuterol, both inhalers and nebulizers, are available in the event of bronchospasm. If a subject 
has an event that is does not immediately respond to care in the research lab, he/she will be transported 
via EMS to UNC Healthcare for treatment. 
10.4. 
Safety Monitoring 
[Discuss how, in what time frame, and by whom subject safety will be monitored and the real-time plan for 
responding to adverse reactions, injury, and new safety concerns. Include criteria to be used for early study 
stopping. In addition to real-time evaluation, include consideration for a DSMB, Safety Monitoring 
Committee, or Independent Medical Monitor, as applicable, and describe in detail what their obligations will 
be. For a multi-site study, describe centralized safety oversight of the sites and communication of safety and 
quality issues] 
Medical Emergency procedures: There is a study physician available for all inhalation procedures. A 
code cart and Automated External Defibrillator (AED) are available in the event of a cardiopulmonary 
event. Albuterol, both inhalers and nebulizers, are available in the event of bronchospasm. If a subject 
has an event that is does not immediately respond to care in the research lab, he/she will be transported 
via EMS to UNC Healthcare for treatment.  
Data Safety Monitoring Plan: Data is initially recorded on paper documents, and the documents are 
maintained in a binder which is kept in a locked research office of study staff. This data is then entered 
into REDCap by an initial data entry person, and confirmed by a second user. 
 
10.5. 
Study Suspension / Early Termination of the Study 
[Describe circumstances that may warrant suspension/termination, e.g., unexpected, significant or 
unacceptable risk to participants; incomplete or unevaluable data; determination of futility. Provide 
actionable study stopping rules.]   
Criteria for safety of a given individual during the study which would suspend the individual from 
further participation in the study will include:  
1. A 20% or greater decline in FEV1 from baseline  
2. Symptoms of respiratory distress.  
3. Heart rate, blood pressure, or respiratory rate increase of more than 30% from baseline values 
(established on the same day).  
4. SpO2 < 90% for > 1 hour. 
 
11. Supporting Documentation and Operational 
Considerations 
 
11.1. 
Regulatory, Ethical, and Study Oversight Considerations 
 

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11.1.1. 
Informed Consent Process 
 
11.1.1.1. Consent/Assent and Other Informational Documents Provided 
to Participants 
 
Consent forms describing in detail the study intervention, study procedures, and risks are given 
to the participant and written documentation of informed consent is required prior to starting 
intervention/administering study intervention.  The following consent materials are submitted 
with this protocol: 
  
1. Consent and Assent form for research study 
11.1.1.2. Consent Procedures and Documentation 
Describe the procedure that will be used to obtain informed consent/HIPAA 
authorization and assent (if applicable): Initial contact with potential subjects will be 
phone or virtual meeting. The study will be described in detail to the subject, 
including why the study is being conducted, the medication being studied, the risks 
and benefits, and what is expected of the subject. Briefly, the subject will be informed 
that they will undergo a screening exam and lung function testing. Subject will inhale 
a radiolabel substance and undergo medical imaging to measure the clearance of 
mucus from the airways. Albuterol will be administered and lung function testing will 
be performed. The imaging will be repeated two more times. The study will be 
completed in one day and will last 6 hours. The major risk of the study is exposure 
from the imaging and inhaling the radiolabeled substance. Radiation history will be 
collected and if it is more than the allowable safety limit, the subject will be excluded. 
The major benefit is to understand how subjects with PCD clear mucus. 
The subject will be given adequate time to read the consent, and consent will be 
obtained prior to any study procedures. Consent may take place on a separate day 
from the baseline procedures. Due to COVID-19, consent forms will be reviewed by 
phone or virtual visit prior to the day of procedure. Subject will sign on the day of the 
procedure.  
For subjects between the age of 12-17 years old, an assent form will be provided with 
age appropriate language. A parent will need to sign a consent form for the subjects.  
Who will obtain consent: Consent will be obtained by a study coordinator or a study 
physician. 
 

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Where will consent process take place: The consent process and review will take 
place over the phone or virtual visit due to COVID-19 precautions and limiting in 
person interactions. Patient will sign the consent form on the day of the study. This 
study will take place at the UNC Meadowmont/Eastowne location. If necessary, the 
UNC CEMALB, located in the Human Studies Facility of the US EPA on Mason Farm 
Road in Chapel Hill will serve as an alternate location.  
How will investigator assure that subjects comprehend the nature of the study, 
procedures, the risks and benefits: The subject will be encouraged to ask questions 
regarding the study and procedures. Open ended questions will be asked of the 
subject to solicit correct responses, to help ensure that the subject understands the 
study commitment, procedures and risks and benefits 
 
 
11.2. 
Study Discontinuation and Closure 
 
This study may be temporarily suspended or prematurely terminated if there is sufficient reasonable 
cause.  Written notification, documenting the reason for study suspension or termination, will be 
provided by the suspending or terminating party to <study participants, investigator, funding agency.  If 
the study is prematurely terminated or suspended, the Principal Investigator (PI) will promptly inform 
study participants, the Institutional Review Board (IRB), and sponsor and will provide the reason(s) for 
the termination or suspension.  Study participants will be contacted, as applicable, and be informed of 
changes to study visit schedule. 
  
Circumstances that may warrant termination or suspension include, but are not limited to: 
• 
Determination of unexpected, significant, or unacceptable risk to participants  
• 
Demonstration of efficacy that would warrant stopping    
• 
Insufficient compliance to protocol requirements 
• 
Data that are not sufficiently complete and/or evaluable 
• 
Determination that the primary endpoint has been met 
• 
Determination of futility 
 
Study may resume once concerns about safety, protocol compliance, and data quality are addressed, 
and satisfy the sponsor, IRB and/or Food and Drug Administration (FDA).] 
 
 
No adverse events are expected from the study procedures. Criteria for safety of a given individual 
which would suspend the individual from further participation in the study will include:  
1. A 20% or greater decline in FEV1 from baseline  
2. Symptoms of respiratory distress.  
3. Heart rate, blood pressure, or respiratory rate increase of more than 30% from baseline values 
(established on the same day).  
4. SpO2 < 90% for > 1 hour.  
 

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Criteria for safety within the entire protocol (failure of which would result in suspension of further 
study until consultation with the IRB) will include the following: 
 1. If 2 of the first 10 participants fail the individual safety criteria outlined above, the entire protocol will 
be suspended pending review by the IRB.  
2. No occurrence of any Serious Adverse Event  
 
Subjects will be under direct supervision throughout MCC measurements and will either have contact 
information for after-hours access to a study physician or the UNC Health 24 hour line. 
Screen failure procedures. For subjects who do not meet enrollment criteria, all study related activity 
will stop once it is determined that the subject does not meet criteria. 
 
11.3. 
Confidentiality and Privacy 
 
Participant confidentiality and privacy is strictly held in trust by the participating investigators, their 
staff, and the sponsor(s) and their interventions. This confidentiality is extended to cover testing of 
biological samples and genetic tests in addition to the clinical information relating to participants. 
Therefore, the study protocol, documentation, data, and all other information generated will be held in 
strict confidence. No information concerning the study or the data will be released to any unauthorized 
third party without prior written approval of the sponsor.  
 
All research activities will be conducted in as private a setting as possible. The study participant’s 
contact information will be securely stored at each clinical site for internal use during the study. At the 
end of the study, all records will continue to be kept in a secure location for as long a period as dictated 
by the reviewing IRB, Institutional policies, or sponsor requirements. 
 
 
Certificate of Confidentiality (if applicable) 
 
To further protect the privacy of study participants, a Certificate of Confidentiality will be issued by the 
National Institutes of Health (NIH).  This certificate protects identifiable research information from 
forced disclosure. It allows the investigator and others who have access to research records to refuse to 
disclose identifying information on research participation in any civil, criminal, administrative, legislative, 
or other proceeding, whether at the federal, state, or local level. By protecting researchers and 
institutions from being compelled to disclose information that would identify research participants, 
Certificates of Confidentiality help achieve the research objectives and promote participation in studies 
by helping assure confidentiality and privacy to participants. 
 
 
11.4. 
Future Use of Stored Specimens and Data 
No specimens will be collected as part of this study.  
We may use de-identified data from this study in future research without additional consent. 
 

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11.5. 
Key Roles and Study Governance 
Principal Investigator 
Medical Monitor 
Lawrence E. Ostrowski, PhD 
Scott H. Donaldson, MD 
University of North Carolina 
University of North Carolina 
Address 
6123 Thurston Bowles Bldg, 
Chapel Hill, NC 27599 
2204 Marsico Hall, Chapel Hill, 
NC, 27599 
919-843-7177 
919-966-8980 
ostro@medu.unc.edu 
Scott_donaldson@med.unc.edu 
 
 
11.6. 
Safety Oversight 
 
Data Safety Monitoring Plan: Data is initially recorded on paper documents, and the documents are 
maintained in a binder which is kept in a locked office within the Meadowmont/Eastowne or EPA 
Human Studies facility. Data will be stored in a secure facility with a guarded entrance and requires 
identification for entry. This data is then entered into REDCap by an initial data entry person and 
confirmed by a second user. 
Medical Monitor 
Scott H. Donaldson, MD, has agreed to serve as an independent medical monitor (MM) for the human 
subjects studies associated with Dr. Ostrowski’s RO1, titled "Functional Studies of Novel Genes Mutated 
in Primary Ciliary Dyskinesia II: Genotype to Phenotype".  Dr. Donaldson is a Professor of Medicine, 
Division of Pulmonary Diseases and Critical Care Medicine, and the Associate Director of the Marsico 
Clinical and Translational Research Center, has experience with multiple clinical trials investigating 
mucociliary clearance in cystic fibrosis, and so is ideally suited to provide safety oversight for this 
project. 
In regards to this proposal, the independent MM has the responsibility to 1) review and evaluate 
information relevant to the safety of the study.  Specifically, this involves reviewing the protocols to be 
followed, including protocol halting rules, and addressing any safety concerns. 2)  The MM will advise 
the protocol team on safety oversight. 3) The MM will be also be responsible for monitoring, collecting 
and reporting data related to unanticipated problems and adverse events.  
In practice, the MM will review the protocols with the appropriate investigators before the studies 
begin. The MM will be notified of any unanticipated problems and adverse events immediately, and will 
report SAEs to the UNC IRB and to the NHLBI program officer within 48 hours. In the absence of any 
unanticipated problems and adverse events, the MM will review the protocol and results of the studies 
on a regular basis. Because of the irregular timing of subject recruitment, this may occur more 
frequently, but at least once yearly and after every 6 subjects complete the study. 

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11.7. 
Clinical Monitoring 
 
N/A 
 
 
11.8. 
Quality Assurance and Quality Control  
 
N/A 
 
11.9. 
Data Handling and Record Keeping 
11.9.1. 
Data Collection and Management Responsibilities 
Monitoring Plan: We will use REDCap for data management. The data will be entered by one person 
(typically the coordinator or, for lab analysis, a lab staff member) and then checked by a second person. The 
data is not marked "complete" until the 2nd person verifies the entry. REDCap creates a data dictionary when 
the database is established, this is used as the codebook.  
Database documentation: All databases using equipment generated have a date and user attached to the 
electronic file. Data will be linked with a codebook (i.e. sample ID) using Excel. A second entry is done 
manually in a lab notebook, which includes date, samples assayed, assay used, any issues (like missing 
samples/data), reference to where data is stored, and a printout of the data (all as hardcopies in a lab 
notebook). Adherence to the codebook is ensured by having a second individual to check the entries. The PI 
takes responsibility for data management computations.  
Case report forms: Case report forms will be developed by the study team, using templates from previous 
studies. These forms are maintained by study coordinators and reviewed by investigators as needed.  
How will confidentiality be maintained: Subjects will be issued a subject number when they enroll into the 
study, and this number will only be used to label samples. All PHI will be maintained by the study coordinator 
or the investigators and will be kept in locked areas when not in use. The identifiers that go into REDCap will 
only be accessible to those who need them for their job, specifically study physicians, investigators and 
coordinators. Subjects sometimes undergo procedures – such as sitting in front of the gamma camera – at 
the same time, however other than basic introductions, nothing about one subject is disclosed to the other 
subject. 
11.9.2. 
Study Records Retention 
 
Study documents will be retained for a period of 3 years from the dates of Federal Financial Report (FFR) 
Submission.  
 

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11.10. Protocol Deviations 
A protocol deviation is any noncompliance with the clinical trial protocol, International Conference on 
Harmonisation Good Clinical Practice (ICH GCP). The noncompliance may be either on the part of the 
participant, the investigator, or the study site staff. Protocol deviations are not permitted. Protocol 
deviations must be sent to the reviewing Institutional Review Board (IRB) per their policies. The site 
investigator is responsible for knowing and adhering to the reviewing IRB requirements. 
 
11.11. Publication and Data Sharing Policy  
 
This study will be conducted in accordance with the following publication and data sharing policies and 
regulations: 
National Institutes of Health (NIH) Public Access Policy, which ensures that the public has access to the 
published results of NIH funded research. It requires scientists to submit final peer-reviewed journal 
manuscripts that arise from NIH funds to the digital archive PubMed Central upon acceptance for 
publication. 
 
11.12. Conflict of Interest Policy 
N/A 
 
12. Additional Considerations 
 
N/A 
 
13. References 
 
1. Adam J. Shapiro, Stephanie D. Davis, Deepika Polineni, Michael R. Knowles, Margaret W. Leigh, 
and Valery Lavergne, et al. Diagnosis of Primary Ciliary Dyskinesia: An Official ATS Clinical 
Practice Guideline: Executive Summary. Am J Respir Crit Care Med. 2018; 197: 1524–1533. 
2. Wanner A, Salathe M, O'Riordan TG. Mucociliary clearance in the airways. Am J Respir Crit Care 
Med. 1996;154:1868–1902. 
3. Lobo J, Zariwala MA, Noone PG. Primary ciliary dyskinesia. Semin Respir Crit Care Med. 
2015;36(2):169-179. doi:10.1055/s-0035-1546748 

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4. Knowles MR, Ostrowski LE, Leigh MW, Sears PR, Davis SD, Wolf WE, et al. Mutations in RSPH1 
cause primary ciliary dyskinesia with a unique clinical and ciliary phenotype. Am J Respir Crit Care 
Med. 2014;189:707–717. 
5. Frommer A, Hjeij R, Loges NT, Edelbusch C, Jahnke C, Raidt J, et al. Immunofluorescence analysis 
and diagnosis of primary ciliary dyskinesia with radial spoke defects. Am J Respir Cell Mol Biol. 
2015;53:563–573. 
6. David SD, Rosenfeld M, Lee HS, Ferkol TW, Sagel SD, Dell SD, Milla C, Pittman JE, Shapiro AJ, 
Sullivan KM, Nykamp KR, Krischer JP, Zariwala MA, Knowles MR, Leigh MW. Primary Ciliary 
Dyskinesia: Longitudinal Study of Lung Disease by Ultrastructure Defect and Genotype Am J Respir 
Crit Care Med. 2019. 
7. Laube BL, Karmazyn YJ, Orens JB, Mogayzel PJ Jr. Albuterol improves impaired mucociliary 
clearance after lung transplantation. J Heart Lung Transplant. 2007 Feb;26(2):138-44. doi: 
10.1016/j.healun.2006.11.005. PMID: 17258147. 
8. Tamoki J, Chiyotani A, Sakai N, Konno K. Simulation of ciliary motility mediated by atypical B-
adrenoceptor in canine bronchial epithelium. Life Sci. 1993; 53; 1509-1515.  
9. Devalia JL, Sapsford RJ, Rusznak C, Toumbis MJ, Davies RJ. The effects of salmeterol and 
salbutamol on ciliary beat frequency of cultured human bronchial epithelial cells, in vitro. Pulm 
Pharmacol. 1992; 5: 257-263. 
10. Bennett WD, Wu J, Fuller F, Balcazar JR, Zeman KL, Duckworth H, Donn KH, O'Riordan TG, 
Boucher RC, Donaldson SH. Duration of action of hypertonic saline on mucociliary clearance in the 
normal lung. Journal of applied physiology 2015; 118: 1483-1490.   – methods reference 
11. Noone PG, Leigh MW, Sannuti A, Minnix SL, Carson JL, Hazucha M, Zariwala MA, Knowles MR. 
Primary ciliary dyskinesia: diagnostic and phenotypic features. Am J Respir Crit Care Med. 2004 Feb 
15;169(4):459-67. doi: 10.1164/rccm.200303-365OC. Epub 2003 Dec 4. PMID: 14656747. 
12. Mosquera RA, Nielsen KG, Gardner L, Holgersen, MG, Hogg C, Carr S, Richardson, Olm M, 
Athanazio RA, Rached S, Castillo DJ,  et al. Prevalence of Reversible Airway Obstruction for Specific 
Ultrastructural Defects in Primary Ciliary Dyskinesia. Am J Respir Crit Care Med 2020;201:A5344. 
13. Yin W, Livraghi-Butrico A, Sears PR, Rogers TD, Burns KA, Grubb BR, Ostrowski LE. Mice with a 
Deletion of Rsph1 Exhibit a Low Level of Mucociliary Clearance and Develop a Primary Ciliary 
Dyskinesia Phenotype. Am J Respir Cell Mol Biol. 2019 Sep;61(3):312-321. doi: 10.1165/rcmb.2017-
0387OC. PMID: 30896965; PMCID: PMC6839924. 
14. Ostrowski LE, Yin W, Rogers TD, Busalacchi KB, Chua M, O'Neal WK, Grubb BR. Conditional 
deletion of dnaic1 in a murine model of primary ciliary dyskinesia causes chronic rhinosinusitis. Am J 
Respir Cell Mol Biol. 2010 Jul;43(1):55-63. doi: 10.1165/rcmb.2009-0118OC. Epub 2009 Aug 12. 
PMID: 19675306; PMCID: PMC2911571. 
15. Bennett WD, Laube BL, Corcoran T, Zeman K, Sharpless G, Thomas K, Wu J, Mogayzel PJ Jr, 
Pilewski J, Donaldson S. Multisite comparison of mucociliary and cough clearance measures using 
standardized methods. J Aerosol Med Pulm Drug Deliv. 2013 Jun;26(3):157-64. doi: 
10.1089/jamp.2011.0909. Epub 2013 Mar 21. PMID: 23517172. 
16. Zeman KL, Wu J, Donaldson SH, Bennett WD. Comparison of 133 xenon ventilation equilibrium 
scan (XV) and 99m technetium transmission (TT) scan for use in regional lung analysis by 2D gamma 
scintigraphy in healthy and cystic fibrosis lungs. J Aerosol Med Pulm Drug Deliv. 2013;26(2):94-100. 
doi:10.1089/jamp.2012.0982 
17. Zeman KL, Wu J, Bennett WD. Targeting aerosolized drugs to the conducting airways using very 
large particles and extremely slow inhalations. J Aerosol Med Pulm Drug Deliv. 2010 Dec;23(6):363-
9. doi: 10.1089/jamp.2008.0711. Epub 2010 Sep 23. PMID: 20863250. 

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14. List of Appendices 
This section should include a listing of any tables, questionnaires, investigator brochure, device manual, 
subject ‘handouts’, etc that should accompany the study protocol.   
 
Appendix 1: Adult Consent Form 
 
Appendix 2: COVID-19 Screening Hand-out: Questions which will be used for COVID-19 
screening.   
 
Appendix 3: Safety Outcome Screening: Questions to screen any changes in health status 
before and during the study  
 
 

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Appendix 1: Adult Consent Form 
 
University of North Carolina at Chapel Hill 
Consent to Participate in a Research Study 
Adult Participants 
 
Consent Form Version Date: 02/08/2023 
IRB Study # 20-3465 
Title of Study: Functional Studies of Novel Genes Mutated in Primary Ciliary Dyskinesia 
II: Genotype to Phenotype 
Principal Investigator: Lawrence Ostrowski, PhD 
Principal Investigator Department: Pediatric Pulmonary Division 
Principal Investigator Phone number: 919-843-7177 
Principal Investigator Email Address: ostro@med.unc.edu 
Funding Source and/or Sponsor: NIH 
Study Contact Telephone Number: 919-962-9841 
Study Contact Email: corinne.lawler@unc.edu  
This is a consent form. It provides a summary of the information the research team will discuss 
with you.  If you decide that you would like to take part in this research study, you would sign 
and date this form to confirm your decision.  If you sign and date this form, you will receive a 
signed and dated copy of this form for your records.   
CONCISE SUMMARY 
The purpose of this study is to determine how patients with primary ciliary dyskinesia (PCD) 
clear mucus differently based on their genetic mutation, and to determine if albuterol can help 
them clear mucus from their airways better.  
Participants will undergo screening with basic physical exam and lung function testing. 
Participants will then inhale a radiolabeled substance and undergo medical imaging to measure 
the clearance of mucus in the airways. Albuterol will be administered after lung function testing 
will be repeated. Finally, imaging will be repeated two more times. The study will be completed 
in one day and will last about 6 hours. 
The major risk of the study is radiation exposure from the inhaled radiolabel substance. The 
radiation exposure is very small, but radiation history will be collected and if it is more than the 
safety limits, then you will not be able to participate in the study. The major benefit of the study 
is to try to understand how patients with PCD clear mucus and if albuterol treatment will help.  
You should not join this research study until all of your questions are answered. 

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If you are interested in learning more about the study, please continue reading below.   
What are some general things you should know about research studies? 
You are being asked to take part in a research study.  To join the study is voluntary. 
You may choose not to participate, or you may withdraw your consent to be in the study, for any 
reason, without penalty. 
Things to know before deciding to take part in a research study:  
o The main goal of a research study is to learn things to help patients in the future. 
o The main goal of regular medical care is to help each patient.  
o The decision to join or not join the research study will not cause you to lose any medical 
benefits.  If you decide not to take part in this study, your doctor will continue to treat 
you.  
o Your medical records may become part of the research record.  If that happens, your 
medical records may be looked at and/or copied by the sponsor of this study and 
government agencies or other groups associated with the study. 
Deciding not to be in the study or leaving the study before it is done will not affect your 
relationship with the researcher, your health care provider, or the University of North Carolina-
Chapel Hill. If you are a patient with an illness, you do not have to be in the research study in 
order to receive health care. 
Details about this study are discussed below.  It is important that you understand this information 
so that you can make an informed choice about being in this research study.  
 
You will be given a copy of this consent form.  You should ask the researchers named above, or 
staff members who may assist them, any questions you have about this study at any time. 
 
What is the purpose of this study? 
The purpose of this research study is to compare how patients with primary ciliary dyskinesia 
with different genetic mutations clear mucus from your lungs differently. It will also test whether 
albuterol, an inhaled medication, can help clear mucus better.  
Are there any reasons you should not be in this study? 
You should not be in this study if you: 
1. Are pregnant or nursing a child. 
2. Take medications in the beta blocker family (labetalol, propranolol, etc.) 
3. Smoke or vape tobacco products 
How many people will take part in this study? 
Approximately 32 people who are 12 years of age or older at this institution will take part in this 
study. 
 
How long will your part in this study last? 

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The study will approximately 8 hours.  However, you will also receive screening and reminder 
phone calls in the days leading up to the study.  
 
How will you be contacted during the study? 
In addition to phone calls, the study team would like to communicate with you by text message 
and e-mail, however, you may say “no” to receiving these messages and still participate in this 
study. If you say “yes”, messages may contain personal information about you and may be sent 
or received by the study team’s personal electronic devices or in a method that is not be able to 
be encrypted (protected) and there is the risk your information could be shared beyond you and 
the study team. This information may include study visit reminders and notifications to contact 
the study team. Communication via text message or e-mail will only be used with adult 
participants or parent/legal guardian(s) of minors. 
If you wish to stop receiving unprotected communication from the study team or have lost access 
to your device, please notify the study team using the study contact information on the first page 
of this consent form.  After the study is complete and all research activities finished, or you 
withdraw from the study or request to stop receiving unprotected communication, you will no 
longer receive un-encrypted (un-protected) messages specific to this study. 
_____ Yes, I consent to the study team utilizing the following email to send communication.   
            List Email:                                                           
_____ Yes, I consent to the study team utilizing the following cell phone to send text message 
communication. List Cell Phone:                                                    
_____ No, I do not consent to receive un-protected communication from the study team.    
 
What will happen if you take part in the study? 
Informed Consent: We will explain the study to you. If you decide to participate you will be 
asked to review, sign, and date the consent and HIPAA authorization forms.  
Medical History: Your medical chart will be reviewed and you will be asked questions about 
your health and Primary Ciliary Dyskinesia including current and past illnesses, and use of 
medications.  
Demographic Information: We will collect information about you including, sex, birthdate and 
race.  
Physical Examination: Your study doctor will listen to your lungs and heart. They will briefly 
examine nose, mouth and neck for any abnormal signs. 
Pregnancy Test: Pregnancy testing is required for participation in this study; all girls and 
women age 12 and older will be tested for pregnancy. Only those testing negative will be 
allowed to participate.  

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Spirometry: This test measures how much air your lungs can hold and how fast you can breathe 
out. You will take a deep breath and then blow into a mouthpiece as hard as you can and for as 
long as you can. You might have to wear soft nose clips during the test to stop air from escaping 
through your nose.  You will be asked to repeat this test at least 3 times.  
Mucociliary Clearance Scan: This procedure is a scan or image of your lungs to see how 
mucus is being cleared. We will first place a source of Cobalt 57 in front of your chest for up to 5 
minutes to obtain a transmission scan (picture) of your lungs. This scan will identify various 
regions of your lungs to help the researchers analyze data from all the gamma camera scans. You 
will then have a background scan of your lungs that lasts 15 minutes. Immediately after this scan 
you will inhale aerosolized radiolabeled solution that will help us see where mucus goes and how 
it moves in the lungs and airways. This procedure lasts about 5 minutes. You will then have 
scans of your lungs during the next 6 hours. You will inhale 4 puffs of albuterol using an inhaler 
and take another scan (picture) of your lungs to see how it affect mucus moving in lungs and 
airways.  After the 2 hour scan you will perform spirometry (lung function test) and 30 voluntary 
coughs over the next 30 minutes of scanning.  
Pre-Visit Reminders: We will provide reminders before your study visits. We will also remind 
you about stopping certain medications 12 hours before the study if applicable.  
Current Medications: You will be asked about all medications and non-drug therapies you are 
currently taking.   
Medication Restrictions: Please inform your study doctor of any medications and non-drug 
therapies that you are currently taking. Your study doctor will tell you whether you can continue 
using a particular medication or non-drug therapy while you are participating in this study.  You 
will be asked not to take the following medications while you are participating in the study:  
- 
Albuterol Inhaler/MDI to be stopped 12 hours before study 
- 
Inhalers which have salmeterol, formoterol or other medicines in the same family to 
be stopped 12 hours before the study 
- 
Airway Clearance Medications such as Hypertonic Saline or Pulmozyme to be 
stopped 24 hours before the study.  
 
Explanation of Study Visit 
Procedures 
 
General Screen (phone) 
Study Visit 
Informed Consent 
 
X 
X 
Medical History, med review 
 
X 
X 
Medications  
 
X 
X 
COVID-19 Screen 
 
X 
X 
Physical Exam 
Complete 
 
X 

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Symptom-Directed 
 
X 
Vital Signs 
 
X 
Urine Pregnancy Test 
 
 
x 
Spirometry 
 
 
x 
Mucociliary Clearance Measurement 
 
 
x 
Approximate Length of Time 
 
~1 hour 
~6 hours 
 
What are the possible benefits from being in this study? 
Research is designed to benefit society by gaining new knowledge.  You will not benefit 
personally from being in this research study.  
 
What are the possible risks or discomforts involved from being in this study? 
There are some minimal risks related to the procedures used in the study. Some of these 
procedures you may have had at your regular clinic visits. There may be uncommon or 
previously unknown risks. You should report any problems to the researcher. 
 
Procedure 
Possible Risk Associated with Procedure 
Spirometry 
There is a small risk of wheezing, shortness of 
breath and lightheadedness. 
Mucociliary Clearance Measurement (Gamma 
scintigraphy and radiolabeled compound) 
The mucociliary clearance scan has radiation 
exposure risks and is estimated to be 44 
mRems. For context, adults in Chapel Hill 
area receive about 300 mRems per year in 
natural radiation exposure.  More information 
on radiation risks are below. 
Albuterol 
Common risks are headache, dizziness, fast 
heartbeat, sore throat, runny nose Less 
common risks are change in blood pressure 
and other cardiovascular changes. There is a 
rare risk of paradoxical bronchospasm 
(airway spasms) and anaphylaxis (allergic 
reaction) that may be life threatening. 
 
Confidentiality:  
Being in any research study has the risk of loss of privacy or confidentiality. Below tells you 
how your information will be protected. 
 
Risks associated with radiation exposure: 
This research study involves exposure to radiation from inhaled radiolabeled sulfur colloid for 
gamma scintigraphy scans. While radiolabeled sulfur colloid is FDA approved for oral and IV 
administration, its use by inhalation is experimental. There is, however, no significant increased 
radiation risk for our experimental vs. FDA-approved use.  Please note that this radiation 
exposure is not necessary for your medical care and is for research purposes only. Since radiation 

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can be especially harmful to a developing fetus, it is important that pregnancy be avoided during 
this study by using effective birth control measures (either hormonal contraceptives, like birth 
control pills; or a barrier method, like condoms. 
 
The radiation dose you receive will be 44mRems. The risk from the radiation dose received from 
this procedure is too small to be detected. The radiation exposure described here is what you will 
get from this research study only. It does not include any exposure you may have received or will 
receive from other tests outside this study that are a part of your medical care.  
 
The average person in the United States receives a radiation exposure of 0.3 rem (or 300 mRem) 
per year from natural background sources, such as the sun, and from radioactive materials that 
are found naturally in the earth’s air and soil. The dose that you will receive from participation in 
this research study (44 mRem) is less than amount you receive from these natural sources in one 
year (equivalent to 15% of exposure from background radiation in one year). 
 
The amount of radiation you will receive in this study has a minimal risk and is below the dose 
guideline established by The University of North Carolina Radiation Safety Committee for 
research subjects. You must inform one of the investigators if you have had any x-rays or other 
radiation exposure within the past year so that we do not exceed the yearly dose limits.  If you 
wish, Dr. Bennett will provide you with additional information and answer any questions you 
may have.  If desired, additional information can be obtained from Marija Ivanovic, Ph.D, 
Chairman of the Radiation Safety Subcommittee of UNC Hospitals at 984-974-7779. 
 
What are the risks to a pregnancy or to a nursing child. 
The risks to pregnancy or a nursing child are unknown. If you are a woman and are pregnant or 
nursing a child, you should not be in the study. 
 
If you choose not to be in the study, what other treatment options do you have? 
There is no treatment in this study.  Your alternative is not to be in this study and continue your 
normal care. 
 
What if we learn about new findings or information during the study?  
You will be given any new information gained during the course of the study that might affect 
your willingness to continue your participation.  
 
The imaging we are using in this research study is not the same quality as imaging that you may 
have as part of your health care. The images will not be reviewed by a doctor who normally 
reads such images (such as a radiologist). As a result, you may not be informed of any 
unexpected findings. The results will not be placed in your medical record.  Occasionally the 
technologist or principal investigator may notice something abnormal on the imaging.  If this 
does occur, the images will be reviewed by a qualified doctor to determine if there is anything of 
clinical importance. If something is found to be important then you, and/or your primary care 
provider will be notified.  Any further follow up and costs associated with the incidental finding 
will be your responsibility. There may be benefits to learning such results (such as early 
detection and treatment of a medical condition), but there are risks as well (such as problems 

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with getting insurance or a job, or feeling worried about a finding for which no treatment is 
required or appropriate). 
  
Do you wish to be informed in case of clinical/relevant unexpected findings?  Please initial in the 
box below if you do not wish to be notified of clinical/relevant unexpected findings.  If you do 
not initial in the box, you will be notified of any findings. 
  
 ______ I do not wish to be notified. 
 
Will I receive any other clinical results?  
Other clinically relevant results of this research will be communicated with you: 
- 
Lung Function testing 
 
How will information about you be protected? 
If you take part in this study, we will make every effort to keep your information confidential.    
We will store all of your research records in locked cabinets and secure computer files. We will 
not put your name on any research data.  Instead, we will label your information or samples with 
a study number.  The master list that links a person’s name to their study number is stored in a 
locked cabinet or on a secure computer file.  
Absolute confidentiality cannot be guaranteed because of the need to give information to these 
parties.  If results of this research are published, we would not use information that identifies 
you.  We would only use your information for research.  
 
These are some reasons that we may need to share the information you give us with others:  
• If it’s required by law.  
• If we think you or someone else could be harmed.  
• Sponsors, government agencies or research staffs sometimes look at forms like this and 
other study records.  They do this to make sure the research is done safely and legally. Anyone 
who reviews study records would keep your information confidential. Agencies or sponsors that 
may look at study records include: 
 National Institute of Health (NIH) 
 Institutional Review Board (IRB) and others responsible for watching over the 
safety, effectiveness, and conduct of the research. 
 
Participants will not be identified in any report or publication about this study.  We may use de-
identified data and/or specimens from this study in future research without additional consent. 
 
What is a Certificate of Confidentiality?  
This research is covered by a Certificate of Confidentiality. With this Certificate, the researchers 
may not disclose or use information, documents or biospecimens that may identify you in any 
federal, state, or local civil, criminal, administrative, legislative, or other proceedings in the 
United States, for example, if there is a court subpoena, unless you have consented for this use. 
 
The Certificate cannot be used to refuse a request for information from personnel of a federal or 
state agency that is sponsoring the study for auditing or evaluation purposes or for information 

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that must be disclosed in order to meet the requirements of the federal Food and Drug 
Administration (FDA). 
 
The Certificate of Confidentiality will not be used to prevent disclosure as required by federal, 
state, or local law, such as mandatory reporting requirements for child abuse or neglect, disabled 
adult abuse or neglect, communicable diseases, injuries caused by suspected criminal violence, 
cancer diagnosis or benign brain or central nervous system tumors or other mandatory reporting 
requirement under applicable law. The Certificate of Confidentiality will not be used if 
disclosure is for other scientific research, as allowed by federal regulations protecting research 
subjects or for any purpose you have consented to in this informed consent document. 
 
You should understand that a Certificate of Confidentiality does not prevent you from 
voluntarily releasing information about yourself or your involvement in this research. If an 
insurer, employer, or other person obtains your written consent to receive research information, 
then the researchers may not use the Certificate to withhold that information. 
You will be asked to sign a separate form ("HIPAA Authorization") to allow researchers to 
review your medical records.  
 
What will happen if you are injured by this research? 
All research involves a chance that something bad might happen to you.  If you are hurt, become 
sick, or develop a reaction from something that was done as part of this study, the researcher will 
help you get medical care, but the University of North Carolina at Chapel Hill has not set aside 
funds to pay you for any such injuries, illnesses or reactions, or for the related medical care.  Any 
costs for medical expenses will be billed to you or your insurance company.  You may be 
responsible for any co-payments and your insurance may not cover the costs of study related 
injuries. 
 
If you think you have been injured from taking part in this study, call the study physician at the 
phone number provided on this consent form.  They will let you know what you should do.  
 
By signing this form, you do not give up your right to seek payment or other rights if you are 
harmed as a result of being in this study. 
 
What if you want to stop before your part in the study is complete? 
You can stop being part of the study at any time, without penalty.  The investigators also have 
the right to stop your participation at any time. This could be because you have had an 
unexpected reaction, or have failed to follow instructions, or because the entire study has been 
stopped. 
 
If you withdraw or are withdrawn from this study all data collected up until the point of 
withdrawal will be retained, however no additional information will be collected unless you 
provide additional written permission for further data collection at the time of your withdrawal. 
 
Will you receive anything for being in this study? 
You will be receiving $150.00 for taking part in this study. Any payment provided for 
participation in this study may be subject to applicable tax withholding obligations. 

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All study payments will be paid through the University of North Carolina Disbursement Services 
office. Receipt of payment can take up to 4 weeks or longer. Your name, address, and U.S. tax 
payer identification number (SSN or ITIN) are required to process payments and/or to report 
taxable income to the IRS.  You must complete a W-9 (for U.S. persons) or W-8BEN and the 
Foreign Vendor Withholding Assessment with supporting documents (for non-resident aliens) in 
order to receive payment for participation. 
 
U.S. person participants must complete Form W-9 in order to receive payment for 
participation.  If payment by UNC equals or exceeds $600 per calendar year for U.S. persons, 
UNC will report the amount to the Internal Revenue Service on Form 1099.  Nonresident alien 
participants must complete Form W-8BEN and the Foreign Vendor Withholding Assessment 
with supporting documents in order to receive payment for participation.  Payments to 
nonresident alien participants may be subject to tax withholding and are generally reported to the 
Internal Revenue Service on Form 1042-S.  This information will not be linked to any of the 
study data and will only be used for payment purposes. 
 
If you do not provide your SSN or ITIN, or complete the appropriate documentation noted 
above, we cannot issue you a payment for participation.  However, you may still choose to 
participate in this study. 
 
We will also reimburse you for any travel costs needed for the study such as parking, 
transportation, and hotel if applicable.  
Will it cost you anything to be in this study? 
It will not cost you anything to be in this study.  
 
What if you are a UNC student? 
You may choose not to be in the study or to stop being in the study before it is over at any 
time.  This will not affect your class standing or grades at UNC-Chapel Hill.  You will not be 
offered or receive any special consideration if you take part in this research. 
 
What if you are a UNC employee? 
Taking part in this research is not a part of your University duties and refusing will not affect 
your job.  You will not be offered or receive any special job-related consideration if you take part 
in this research. 
 
Who is sponsoring this study? 
This research is funded by National Institutes of Health (NIH).  This means that the research 
team is being paid by the sponsor for doing the study.  The researchers do not, however, have a 
direct financial interest with the sponsor or in the final results of the study. 
 
What if you have questions about this study? 
You have the right to ask, and have answered, any questions you may have about this research. If 
you have questions about the study (including payments), complaints, concerns, or if a research-
related injury occurs, you should contact the researchers listed on the first page of this form.  

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A description of this clinical trial will be available on www.clinicaltrials.gov, as required by U.S. 
Law. This website will not include information that can identify you. At most, the website will 
include a summary of the results. You can search this website at any time. 
 
What if you have questions about your rights as a research participant? 
All research on human volunteers is reviewed by a committee that works to protect your rights 
and welfare.  If you have questions or concerns about your rights as a research subject, or if you 
would like to obtain information or offer input, you may contact the Institutional Review Board 
at 919-966-3113 or by email to IRB_subjects@unc.edu. 
 
 

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Participant’s Agreement: 
 
I have read the information provided above.  I have asked all the questions I have at this time.  I 
voluntarily agree to participate in this research study. 
 
______________________________________________________ 
Signature of Research Participant 
 
____/____/____ 
Date 
 
______________________________________________________ 
Printed Name of Research Participant 
 
Researcher’s Signature  
I have fully explained the research study described by this form. I have answered the participant 
and/or parents’/guardians’ questions and will answer any future questions to the best of my 
ability. I will tell the family and/or the person taking part in this research of any changes in the 
procedures or in the possible harms/possible benefits of the study that may affect their health or 
their willingness to stay in the study.  
__________________________________  
 
 
 
__________________ 
Printed Name of Person Conducting the 
 
 
 
 
Position 
Informed Consent Discussion  
________________________________________   
 
 
____/____/____  
Signature of Person Conducting the   
 
 
 
 
Date 
Informed Consent Discussion 
 
 
 
 
 
 
 
 
 

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Appendix 2: COVID-19 Screening Hand-out: 
 
 
 
 
 

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Appendix 3: Safety Outcome Screening 
 
Symptom Questionnaire:  
 We will use the following symptom questionnaire:  
1. Do you have any health concerns since your last visit?  
2.  Have you needed to see a doctor for any reason since your last visit?  
3.  Have you needed to use any over the counter medications?  
4.  Have you had any specific problems with cough, wheezing, or needed any extra allergy or asthma 
medications?  
5. If the answers to questions 1, 2, 3, or 4 are “yes”, the subject will be assessed by a study physician.