Evaluate Established Anti-DEFA5 mAbs Diagnostic Efficacy and Safety in IBD
Conditions: Inflammatory Bowel Diseases, Ulcerative Colitis, Crohn Colitis, Indeterminate Colitis
Sponsor: Meharry Medical College

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ClinicalTrials.gov PRS DRAFT Receipt (Working Version) 
Last Update: 11/15/2022 20:55 
ClinicalTrials.gov ID: [Not yet assigned] 
  
Study Identification 
Unique Protocol ID: 22-08-1235 
Brief Title: Evaluate Established Anti-DEFA5 mAbs Diagnostic Efficacy and Safety in IBD 
Official Title: Evaluate Established Anti-DEFA5 mAbs Diagnostic Efficacy and Safety in IBD 
Secondary IDs:  
Study Status 
Record Verification: November 2022 
Overall Status: Not yet recruiting 
Study Start: June 1, 2023 [Anticipated] 
Primary Completion: May 31, 2025 [Anticipated] 
Study Completion: June 30, 2025 [Anticipated] 
Sponsor/Collaborators 
Sponsor: Meharry Medical College 
Responsible Party: Principal Investigator 
Investigator: Amosy M’Koma [amkoma] 
Official Title: Professor 
Affiliation: Meharry Medical College 
 
Collaborators:  
Oversight 
U.S. FDA-regulated Drug:  
U.S. FDA-regulated Device:  
U.S. FDA IND/IDE:  
Human Subjects Review: Board Status: Exempt 
Data Monitoring: Yes 
Study Description 
Brief Summary: Investigators propose to validate efficacy and safety of the detection of DEFA5 
in the diagnosis of the colonic IBD using longitudinal vs. cross-sectional studies 
of known patient clinical data to correlate with their endoscopy biopsy data. 
30% of colonic IBD patients cannot be accurately diagnosed (CC vs. UC) in a 
timely manner even when a state-of-the-art classification system of combined 
clinical, endoscopic, radiologic and histologic tools is used. When the diagnostic 
classification for these two diseases is inconclusive, the condition is termed 
indeterminate colitis (IC). Here, the central medical challenge is the 
discrimination of IBD into the specific subtypes with high accuracy, as it greatly 
affects surgical care of patients. Diagnostic accuracy of IC into either authentic 
UC or CC is of utmost importance when determining a patient’s candidacy for 

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RPC-IPAA surgery, the standard curative surgical procedure for UC. Further, 
incorrect diagnosis and treatment carry potential morbidity from inappropriate 
and unnecessary surgery and costs. The success outcomes of RPC-IPAA 
surgery and convalescence depend on correct diagnosis. To address IBD 
diagnosis ambiguity and delays in IBD clinical settings, investigators developed 
a proteomic signature to discriminate between UC and CC patients that also will 
predict the outcome of IC patients for their eventual progress to either UC or 
CC. Our published data has shown robust evidence supporting presence of 
human alpha-defensin 5 (DEFA5) in areas of the colon mucosa with aberrant 
expression of apparent Paneth cell-like cells (PCLCs) or crypt cell-like cells 
(CCLCs), which identifies an area of colonic ileal metaplasia, consistent with the 
diagnosis of CC. DEFA5 bioassay discriminated CC and UC in a cohort of all IC 
patients with accuracy. A fit logistic model with group CC and UC as the 
outcome and the DEFA5 as independent variable differentiator with a positive 
predictive value of 96%. These findings were obtained solely from colectomy 
specimens for both the discovery and validation analyses. Investigators believe 
that use of endoscopy biopsies would be indifferent, which is the purpose of this 
prospective patient centered clinical study. Investigators propose to 
demonstrate that UC and CC, the two unsolved medical subtypes of pathology 
with no drugs for a cure, can accurately be distinguished molecularly by 
examining CCLCs-secreted DEFA5 in colonic endoscopy biopsies instantly. 
Our proposal is highly innovative, as it highlights the robustness of DEFA5 and 
its clinical relevance to IBD is both in science and the anticipated impact, as 
investigators seek to better understand difficulty to determine ‘subtypes” and 
translate that to improve diagnosis, treatment, clinical outcomes, and quality of 
life for patients and the realm of clinical care. DEFA5 immunoreactivity in 
colonic endoscopy biopsies could be a rapid potential diagnostic signature to 
resolve IC into authentic UC and CC with a first clinic endoscopy biopsy. IC is 
likely to be eliminated for good. 
Detailed Description: The clinical relevance of this proposed screen is that it would lead to the elusive 
and accurate diagnosis to circumvent the inexact IC patients into authentic CC 
and UC with a first clinic endoscopy biopsy. The central medical challenge in 
endoscopic medicine and colorectal surgery is the discrimination of colonic IBD 
into the subtypes with high accuracy because it greatly affects surgical care of 
patients. Incorrect diagnosis and surgery carry potential morbidity from 
inappropriate and unnecessary surgery and cost. Our published data has 
shown robust evidence supporting presence of human alpha-defensin 5 
(DEFA5) in areas of the colon mucosa with aberrant expression of apparent 
Paneth cell-like cells (PCLCs) / crypt cell-like cells (CCLCs), which identifies an 
area of colonic ileal metaplasia, consistent with the diagnosis of CC. 
Investigators propose to demonstrate that UC and CC, the two unsolved 
medical subtypes of GI pathology with no drugs for a cure, may accurately be 
distinguished molecularly by examining CCLCs-secreted DEFA5 in colonic 
endoscopy biopsies instantly. Our proposal is highly innovative, as it highlights 
the robustness efficacy and safety of DEFA5 and its clinical relevance in IBD 
diagnostics. The goal of this proposal is to develop a clinical approach to 
circumvent diagnostic ambiguity and delay, as well as permit timely and 
accurate diagnosis of colonic IBD. DEFA5 immunoreactivity in colonic 
endoscopy biopsies could be a potential diagnostic signature that accurately 
diagnoses CC and provides the basis to resolve ambiguity in the diagnosis of 
IBD to not only circumvent diagnostic delay, but also permit timely, accurate 
diagnosis and timely prescription of appropriate treatment options, an 
Affordable, Sensitive, Specific, User-friendly, Robust and Rapid, 
Equipment-free, and Deliverable (ASSURED) bioassay that may delineate 
subtypes of IBD during the first clinic endoscopy biopsy visit. This bioassay is 
specific, sensitive, linear, affordable, low risk, and less invasive. Investigators 
hypothesize that aberrant expression of DEFA5 secreting CCLCs in colonic 
crypt of IBD patients may be exploited as a reliable diagnostic signature to 
highly differentiate CC from UC in otherwise IC patients during the first clinic 
visit endoscopy biopsy without delay. Investigators foresee no issues pertaining 

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to this proposal as an established sampling error by endoscopic biopsy making 
it does not interfere with results and apropos of this project that each biopsy has 
complement CCLCs with co-localized DEFA5 clearly restricted in areas of the 
mucosa with aberrant CCLCs identifies a ubiquitously colonic ileal metaplasia 
that is consistent with the diagnosis of authentic CC. If successful, widespread 
use of this approach would not only revolutionize provide accurate diagnoses 
and the correct treatment regimens for IBD patients, but also it will improve 
health outcomes and patient quality of life, while reducing medical 
complications and/or unnecessary drugs, surgeries, & costs. 
 
Conditions 
Conditions: Inflammatory Bowel Diseases 
Ulcerative Colitis 
Crohn’s Colitis 
Indeterminate Colitis 
 
Keywords: Inflammatory bowel disease 
Crohn’s colitis 
Ulcerative colitis 
Indeterminate colitis 
DEFA5 
Molecular diagnostics 
Colonic biopsies 
Colonic ileal metaplasia 
 
Study Design 
Study Type: Observational 
Observational Study Model: Cohort 
Time Perspective: Cross-Sectional 
Biospecimen Retention: Samples With DNA 
Biospecimen Description: The research outlined is consistent with the Non-human Welfare Act Criteria 
because investigators receive de-identified specimens and data from an IRB 
approved repository, and investigators do not have access to the link by which 
investigators may re-identify the specimens and data with their source. The 
research involves the use of human tissue acquired previously using the 
third-party good faith broker mechanism. Since all identifiers are striped, the 
tissue samples are considered non-human subjects. Each specimen is 
assigned a unique barcode identifier to maintain the patient’s confidentiality. In 
no case is the patient’s name ever revealed to the users. All information 
accompanying the specimen(s) uses the bar code identifier. 
Enrollment: 230 [Anticipated] 
Number of Groups/Cohorts: 5 
Groups and Interventions 
Groups/Cohorts 
Interventions 
Crohn’s colitis 
Correlate patient longitudinal study of clinical data and their biopsy data. 
Investigators will determine DEFA5 levels from endoscopy biopsies from 
known authentic CC patients. 
 
Diagnostic Test: Diagnostic Test 
The challenge is the discrimination of 
IBD into the subtypes with accuracy 
because it affects surgical care of 
patients. Investigators have 
developed a signature discriminator 
between UC&CC that also predicts 
the outcome of IC patients into 

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Groups/Cohorts 
Interventions 
authentic UC/CC. This is a bioassay 
that is specific, sensitive, linear, 
affordable, minimal risk, non-invasive 
and constitutes an inexpensive, 
simple to use, point-of-care test 
format. Our published data have 
shown robust evidence supporting 
presence of DEFA5 in the colon 
crypt mucosa with aberrant 
expression of apparent crypt-cell-like 
cells (CCLCs) in areas identified with 
ectopic colon ileal metaplasia 
consistent with CC. Investigators 
propose to validate that UC/CC, the 
two unsolved medical sub-types of 
pathology can accurately be 
distinguished among IC patients by 
examining CCLCs secreted DEFA5 
levels in endoscopy biopsies 
instantly. Diagnostics relies on the 
expression/localization/activation of 
DEFA5 and the ectopic CCLCs in 
the mucosal crypt of CC patients. 
 
Ulcerative colitis 
Correlate patient longitudinal study of clinical data and their biopsy data. 
Investigators will determine DEFA5 levels from endoscopy biopsies from 
known authentic UC patients. 
 
Diagnostic Test: Diagnostic Test 
The challenge is the discrimination of 
IBD into the subtypes with accuracy 
because it affects surgical care of 
patients. Investigators have 
developed a signature discriminator 
between UC&CC that also predicts 
the outcome of IC patients into 
authentic UC/CC. This is a bioassay 
that is specific, sensitive, linear, 
affordable, minimal risk, non-invasive 
and constitutes an inexpensive, 
simple to use, point-of-care test 
format. Our published data have 
shown robust evidence supporting 
presence of DEFA5 in the colon 
crypt mucosa with aberrant 
expression of apparent crypt-cell-like 
cells (CCLCs) in areas identified with 
ectopic colon ileal metaplasia 
consistent with CC. Investigators 
propose to validate that UC/CC, the 
two unsolved medical sub-types of 
pathology can accurately be 
distinguished among IC patients by 
examining CCLCs secreted DEFA5 
levels in endoscopy biopsies 
instantly. Diagnostics relies on the 
expression/localization/activation of 
DEFA5 and the ectopic CCLCs in 
the mucosal crypt of CC patients. 
 
Indeterminate colitis 
Correlate patient longitudinal study of clinical data and their biopsy data. 
Investigators will determine DEFA5 levels from endoscopy biopsies from 
Diagnostic Test: Diagnostic Test 
The challenge is the discrimination of 
IBD into the subtypes with accuracy 

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Groups/Cohorts 
Interventions 
known IC patients (into authentic UC and CC) 
 
because it affects surgical care of 
patients. Investigators have 
developed a signature discriminator 
between UC&CC that also predicts 
the outcome of IC patients into 
authentic UC/CC. This is a bioassay 
that is specific, sensitive, linear, 
affordable, minimal risk, non-invasive 
and constitutes an inexpensive, 
simple to use, point-of-care test 
format. Our published data have 
shown robust evidence supporting 
presence of DEFA5 in the colon 
crypt mucosa with aberrant 
expression of apparent crypt-cell-like 
cells (CCLCs) in areas identified with 
ectopic colon ileal metaplasia 
consistent with CC. Investigators 
propose to validate that UC/CC, the 
two unsolved medical sub-types of 
pathology can accurately be 
distinguished among IC patients by 
examining CCLCs secreted DEFA5 
levels in endoscopy biopsies 
instantly. Diagnostics relies on the 
expression/localization/activation of 
DEFA5 and the ectopic CCLCs in 
the mucosal crypt of CC patients. 
 
Diverticulitis 
Correlate patient longitudinal study of clinical data and their biopsy data. 
Investigators will determine DEFA5 levels from endoscopy biopsies from 
known diverticulitis. 
 
Diagnostic Test: Diagnostic Test 
The challenge is the discrimination of 
IBD into the subtypes with accuracy 
because it affects surgical care of 
patients. Investigators have 
developed a signature discriminator 
between UC&CC that also predicts 
the outcome of IC patients into 
authentic UC/CC. This is a bioassay 
that is specific, sensitive, linear, 
affordable, minimal risk, non-invasive 
and constitutes an inexpensive, 
simple to use, point-of-care test 
format. Our published data have 
shown robust evidence supporting 
presence of DEFA5 in the colon 
crypt mucosa with aberrant 
expression of apparent crypt-cell-like 
cells (CCLCs) in areas identified with 
ectopic colon ileal metaplasia 
consistent with CC. Investigators 
propose to validate that UC/CC, the 
two unsolved medical sub-types of 
pathology can accurately be 
distinguished among IC patients by 
examining CCLCs secreted DEFA5 
levels in endoscopy biopsies 
instantly. Diagnostics relies on the 
expression/localization/activation of 
DEFA5 and the ectopic CCLCs in 
the mucosal crypt of CC patients. 

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Groups/Cohorts 
Interventions 
 
Ileum 
Positive control 
 
Diagnostic Test: Diagnostic Test 
The challenge is the discrimination of 
IBD into the subtypes with accuracy 
because it affects surgical care of 
patients. Investigators have 
developed a signature discriminator 
between UC&CC that also predicts 
the outcome of IC patients into 
authentic UC/CC. This is a bioassay 
that is specific, sensitive, linear, 
affordable, minimal risk, non-invasive 
and constitutes an inexpensive, 
simple to use, point-of-care test 
format. Our published data have 
shown robust evidence supporting 
presence of DEFA5 in the colon 
crypt mucosa with aberrant 
expression of apparent crypt-cell-like 
cells (CCLCs) in areas identified with 
ectopic colon ileal metaplasia 
consistent with CC. Investigators 
propose to validate that UC/CC, the 
two unsolved medical sub-types of 
pathology can accurately be 
distinguished among IC patients by 
examining CCLCs secreted DEFA5 
levels in endoscopy biopsies 
instantly. Diagnostics relies on the 
expression/localization/activation of 
DEFA5 and the ectopic CCLCs in 
the mucosal crypt of CC patients. 
 
Outcome Measures 
Primary Outcome Measure: 
1. Assesse investigator’s two newly developed anti-DEFA5 monoclonal antibodies for immunohistochemistry and 
sandwich ELISAs, and determines the most effective one or both for bioassay kit for IBD diagnostics. 
[Time Frame: Six months] 
 
Secondary Outcome Measure: 
2.  Assess expression of DEFA5 biomarker in colonic biopsies from known patients with the diagnosis of IBD i.e., 
UC, CC, IC, and none-IBD i.e., diverticulitis, diverticulosis, & ileum as positive control, this will differentiate UC 
vs. CC and IC into authentic UC and CC. 
[Time Frame: 12 months] 
Eligibility 
Study Population: A sample size of 230 subjects is sufficient to detect a clinically significant 
difference of 19% between the PPVs of CC and UC using a one-tailed test of 
proportions between the two groups with 80% statistical power and a 5% level 
of significance. This 19% difference represents 80% probability that subjects in 
the CC group with a positive screening test truly have the disease and 77% 
PPV for subjects in UC group. Investigators will add non-IBD samples, 46 in the 
DV arm and 46 in CP arm to strengthen the results. 
Sampling Method: Probability Sample 

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Minimum Age:  
Maximum Age:  
Sex: All 
Gender Based: No 
Accepts Healthy Volunteers: Yes 
Criteria: Inclusion Criteria: 
There will be no specific recruitment of subjects for this research. Subjects are 
consented, in general, for the collection of tissue and serum for banking 
purposes. Not specifically for this project. Biopsies and serum collections are 
performed routinely as standard of care and no specific procedures are 
performed on the subjects for this grant. 
Exclusion Criteria: 
No children 
Contacts/Locations 
Central Contact Person: Kimberly Thomas, RN, CCRP 
Telephone: 615-327-6735 
Email: kthomas@mmc.edu 
 
Central Contact Backup:  
Study Officials: Amosy E M’Koma, MD, MS, PhD 
Study Principal Investigator 
Meharry Medical College 
 
Locations: United States, Tennessee 
Meharry Medical College 
Nashville, Tennessee, United States, 37208-3501 
Contact
: 
Amosy E M’Koma, MD, MS, PhD 615-327-6796   amkoma@mmc.e
IPDSharing 
Plan to Share IPD: Undecided 
IPD will be obtained (i) directly with study PI or (ii) request via a data repository 
 
References 
Citations: [Study Results] Williams AD, Korolkova OY, Sakwe AM, Geiger TM, James 
SD, Muldoon RL, Herline AJ, Goodwin JS, Izban MG, Washington MK, Smoot 
DT, Ballard BR, Gazouli M, M’Koma AE. Human alpha defensin 5 is a 
candidate biomarker to delineate inflammatory bowel disease. PLoS One. 2017 
Aug 17;12(8):e0179710. doi: 10.1371/journal.pone.0179710. eCollection 2017. 
Erratum in: PLoS One. 2017 Dec 6;12 (12 ):e0189551. PubMed 28817680 
[Study Results] Rana T, Korolkova OY, Rachakonda G, Williams AD, Hawkins 
AT, James SD, Sakwe AM, Hui N, Wang L, Yu C, Goodwin JS, Izban MG, 
Offodile RS, Washington MK, Ballard BR, Smoot DT, Shi XZ, Forbes DS, 
Shanker A, M’Koma AE. Linking bacterial enterotoxins and alpha defensin 5 
expansion in the Crohn’s colitis: A new insight into the etiopathogenetic and 
differentiation triggers driving colonic inflammatory bowel disease. PLoS One. 
2021 Mar 9;16(3):e0246393. doi: 10.1371/journal.pone.0246393. eCollection 

-  Page 8   [DRAFT]  - 
2021. PubMed 33690604 
[Study Results] Ballard BR, M’Koma AE. Gastrointestinal endoscopy biopsy 
derived proteomic patterns predict indeterminate colitis into ulcerative colitis 
and Crohn’s colitis. World J Gastrointest Endosc. 2015 Jun 25;7(7):670-4. doi: 
10.4253/wjge.v7.i7.670. PubMed 26140094 
[Study Results] Korolkova OY, Myers JN, Pellom ST, Wang L, M’Koma AE. 
Characterization of Serum Cytokine Profile in Predominantly Colonic 
Inflammatory Bowel Disease to Delineate Ulcerative and Crohn’s Colitides. Clin 
Med Insights Gastroenterol. 2015 May 6;8:29-44. doi: 10.4137/CGast.S20612. 
eCollection 2015. PubMed 26078592 
[Study Results] M’Koma AE. Diagnosis of inflammatory bowel disease: 
Potential role of molecular biometrics. World J Gastrointest Surg. 2014 Nov 
27;6(11):208-19. doi: 10.4240/wjgs.v6.i11.208. Review. PubMed 25429322 
[Study Results] Seeley EH, Washington MK, Caprioli RM, M’Koma AE. 
Proteomic patterns of colonic mucosal tissues delineate Crohn’s colitis and 
ulcerative colitis. Proteomics Clin Appl. 2013 Aug;7(7-8):541-9. doi: 
10.1002/prca.201200107. Epub 2013 May 8. PubMed 23382084 
[Study Results] M’Koma AE, Seeley EH, Washington MK, Schwartz DA, 
Muldoon RL, Herline AJ, Wise PE, Caprioli RM. Proteomic profiling of mucosal 
and submucosal colonic tissues yields protein signatures that differentiate the 
inflammatory colitides. Inflamm Bowel Dis. 2011 Apr;17(4):875-83. doi: 
10.1002/ibd.21442. Epub 2010 Aug 30. PubMed 20806340 
 
Links:  
Available IPD/Information:  
U.S. National Library of Medicine  |  U.S. National Institutes of Health  |  U.S. Department of Health & Human Services