Transfusion of Biotin-Labeled Red Blood Cells

• Sponsor: Gladwin, Mark, MD (Other)
• Overall Status: Terminated
• CT.gov ID: NCT03364686
• Collaborator: National Heart, Lung, and Blood Institute (NHLBI) (NIH)
• Enrollment: 6
• Location: 1
• Arm: 1
• Actual Duration (Months): 21.4
• Patients Per Site Per Month: 0.3

Study Details

Study Description

Brief Summary

The purpose of this research is to evaluate the impact of genetic and biologic factors in blood donors on red blood cell storage stability after autologous transfusion over the different range of storage period of 5-7 days and 35-42 days in healthy volunteers.

Condition or Disease	Intervention/Treatment	Phase
Healthy Volunteers	Biological: Biotin-Labeled Red Blood Cells	Phase 2

Study Design

• Study Type: Interventional
• Actual Enrollment: 6 participants
• Allocation: N/A
• Intervention Model: Single Group Assignment
• Masking: None (Open Label)
• Primary Purpose: Basic Science
• Official Title: Transfusion of Biotin-Labeled Red Blood Cells for the Evaluation of Genetic Factors That Contribute to Donor Differences in Red Blood Cell Storage and Post-Transfusion Red Blood Cell Recovery
• Actual Study Start Date: Sep 16, 2019
• Actual Primary Completion Date: Mar 2, 2021
• Actual Study Completion Date: Jun 29, 2021

Arms and Interventions

Arm	Intervention/Treatment
Experimental: Biotin-Labeled Red Blood Cells Infusion Each participant will receive 2 transfusions of biotin labeled red blood cells.	Biological: Biotin-Labeled Red Blood Cells; We will collect 500 mL of blood. The blood will be processed and split into two bags and labeled with a naturally occurring vitamin, biotin. The blood will then be re-infused back into the same participant at 2 time points (5-7 days and 35-42 days after storage).

Outcome Measures

Primary Outcome Measures

1. Percentage of Biotin Labeled Red Blood Cells [24 hours, 30 days and 60 days after transfusion]

   This will be determined using enumeration of biotinylated red blood cells obtained by flow cytometry from blood samples obtained at various time points from subjects with specified genetic mutations, compared with control subjects without the specified mutation. Percentage of Biotin Labeled Red Blood Cells is determined by Biotin Labeled Red Blood Cells in circulation at each time point/baseline Biotin Labeled Red Blood Cells in circulation x 100.

Secondary Outcome Measures

1. The Percentage of Storage Hemolysis [5-7 days and 35-42 days following blood donation]

   This will be quantified as percent hemolysis, which accounts for the levels of RBC-derived free hemoglobin in response to cold storage. We will define the associations between the primary endpoints and the quantification of storage hemolysis. Quantification of storage hemolysis is based on this equation: (sample hematocrit x levels of free hemoglobin obtained after centrifugation measured in the supernatant / the total amount of sample hemoglobin before centrifugation) x 100.

2. The Percentage of Red Blood Cell Osmotic Hemolysis [5-7 days and 35-42 days following blood donation]

   This will be quantified by the evaluation of osmotic stress assays. We will define the associations between the primary endpoints and the quantification of osmotic hemolysis. The percent osmotic hemolysis is determined by this equation: (supernatant cell-free hemoglobin of pink test-treated RBCs / total amount of hemoglobin) x 100.

3. The Percentage of Red Blood Cell Oxidative Hemolysis [5-7 days and 35-42 days following blood donation]

   This will be quantified by the evaluation of oxidative stress assays. We will define the associations between the primary endpoints and the quantification of oxidative hemolysis. The percent osmotic hemolysis is determined by this equation: (supernatant cell-free hemoglobin of 2,2'-azobis-2-methyl-propanimidamide, dihydrochloride treated RBCs - supernatant cell-free hemoglobin of untreated red blood cells / total amount of hemoglobin) x 100.

Eligibility Criteria

Criteria

• Ages Eligible for Study: 18 Years and Older
• Sexes Eligible for Study: All
• Accepts Healthy Volunteers: Yes

Inclusion Criteria:

• Ages 18 years and older

• Weight ≥110 lbs

• Hemoglobin ≥ 12.5 g/dL or hematocrit ≥ 38% for women and hemoglobin ≥ 13.0 g/dL or hematocrit ≥ 39% for men.

• Meet criteria for autologous blood donation

Exclusion Criteria:

• Subjects with a past medical history or symptoms of blood dyscrasia, diabetes mellitus, hyperlipidemia, obstructive sleep apnea, renal disease, congestive heart failure, significant cardiac disease and / or known peripheral arterial disease.

• Moderate to severe systemic hypertension (systolic blood pressure >140 mmHg and/or diastolic blood pressure > 95 mmHg

• Systolic blood pressure <100 mmHg and/or diastolic blood pressure < 60 mmHg on the study day.

• Positive Direct Antiglobulin Test

• Consumption of biotin supplements or raw eggs within 30 days

• Treatment with antibiotics in the week before initiating study participation to avoid suppression of erythropoiesis, which may accompany infection.

• Blood loss in the previous 8 weeks due to epistaxis, trauma, hemoptysis, gastrointestinal bleeding, diagnostic phlebotomy (> 30 ml)

• Subjects who report tobacco or marijuana smoking within 6 months of study.

• Cognitively impaired subjects, or institutionalized persons and subjects unable or unwilling to complete written informed consent

• Subjects with a history of blood donation within the last 56 days.

• Use of other investigational drugs/devices within 30 days of screening.

• Subjects taking any medication for the treatment of diabetes including insulin

• Females of childbearing potential who are pregnant or unwilling to undergo pregnancy testing; females with positive pregnancy testing on screening day will be excluded.

• History of prior transfusion reaction to blood products.

• Allergic reaction to biotin

• Donors with naturally occurring antibodies against BioRBCs will be excluded from the study

Contacts and Locations

Locations

	Site	City	State	Country	Postal Code
1	University of Pittsburgh	Pittsburgh	Pennsylvania	United States	15213

Sponsors and Collaborators

• Gladwin, Mark, MD
• National Heart, Lung, and Blood Institute (NHLBI)

Investigators

• Principal Investigator: Darrell Triulzi, MD, University of Pittsburgh

Study Documents (Full-Text)

• Study Protocol and Statistical Analysis Plan - Apr 30, 2021
• Informed Consent Form - May 20, 2021

More Information

Publications

None provided.

• Responsible Party: Darrell Triulzi MD, Professor, University of Pittsburgh
• ClinicalTrials.gov Identifier: NCT03364686
• Other Study ID Numbers: STUDY19020356; 5R01HL098032-09
• First Posted: Dec 6, 2017
• Last Update Posted: Apr 21, 2022
• Last Verified: Mar 1, 2022

• Studies a U.S. FDA-regulated Drug Product: Yes
• Studies a U.S. FDA-regulated Device Product: No
• Keywords provided by Darrell Triulzi MD, Professor, University of Pittsburgh: BioRBCs; Biotin labeled red blood cells
• Additional relevant MeSH terms: Biotin

Study Results

Participant Flow

Arm/Group Title	Biotin-Labeled Red Blood Cells Infusion
Arm/Group Description	Each participant will receive 2 transfusions of biotin labeled red blood cells. Biotin-Labeled Red Blood Cells: We will collect 500 mL of blood. The blood will be processed and split into two bags and labeled with a naturally occurring vitamin, biotin. The blood will then be re-infused back into the same participant at 2 time points (5-7 days and 35-42 days after storage).
Period Title: Overall Study
STARTED	6
COMPLETED	4
NOT COMPLETED	2

Baseline Characteristics

Arm/Group Title	Biotin-Labeled Red Blood Cells Infusion
Arm/Group Description	Each participant will receive 2 transfusions of biotin labeled red blood cells. Biotin-Labeled Red Blood Cells: We will collect 500 mL of blood. The blood will be processed and split into two bags and labeled with a naturally occurring vitamin, biotin. The blood will then be re-infused back into the same participant at 2 time points (5-7 days and 35-42 days after storage).
Overall Participants	6
Age (Count of Participants)
<=18 years	0 0%
Between 18 and 65 years	6 100%
>=65 years	0 0%
Sex: Female, Male (Count of Participants)
Female	3 50%
Male	3 50%
Ethnicity (NIH/OMB) (Count of Participants)
Hispanic or Latino	0 0%
Not Hispanic or Latino	6 100%
Unknown or Not Reported	0 0%
Race (NIH/OMB) (Count of Participants)
American Indian or Alaska Native	0 0%
Asian	0 0%
Native Hawaiian or Other Pacific Islander	0 0%
Black or African American	3 50%
White	3 50%
More than one race	0 0%
Unknown or Not Reported	0 0%

Outcome Measures

1. Primary Outcome

Title	Percentage of Biotin Labeled Red Blood Cells
Description	This will be determined using enumeration of biotinylated red blood cells obtained by flow cytometry from blood samples obtained at various time points from subjects with specified genetic mutations, compared with control subjects without the specified mutation. Percentage of Biotin Labeled Red Blood Cells is determined by Biotin Labeled Red Blood Cells in circulation at each time point/baseline Biotin Labeled Red Blood Cells in circulation x 100.
Time Frame	24 hours, 30 days and 60 days after transfusion

Outcome Measure Data

Analysis Population Description

The blood sample at 24 hours from one of the 4 completed subjects was not collected.

Arm/Group Title	Biotin-Labeled Red Blood Cells Infusion
Arm/Group Description	Each participant will receive 2 transfusions of biotin labeled red blood cells. Biotin-Labeled Red Blood Cells: We will collect 500 mL of blood. The blood will be processed and split into two bags and labeled with a naturally occurring vitamin, biotin. The blood will then be re-infused back into the same participant at 2 time points (5-7 days and 35-42 days after storage).
Measure Participants	3
Percentage of Fresh Biotin Labeled Red Blood Cells at 24 Hours	99.2
Percentage of Fresh Biotin Labeled Red Blood Cells at Day 30	77.2
Percentage of Fresh Biotin Labeled Red Blood Cells at Day 60	54.3
Percentage of Aged Biotin Labeled Red Blood Cells at 24 Hours	99.2
Percentage of Aged Biotin Labeled Red Blood Cells at Day 30	75.9
Percentage of Aged Biotin Labeled Red Blood Cells at Day 60	51.9

2. Secondary Outcome

Title	The Percentage of Storage Hemolysis
Description	This will be quantified as percent hemolysis, which accounts for the levels of RBC-derived free hemoglobin in response to cold storage. We will define the associations between the primary endpoints and the quantification of storage hemolysis. Quantification of storage hemolysis is based on this equation: (sample hematocrit x levels of free hemoglobin obtained after centrifugation measured in the supernatant / the total amount of sample hemoglobin before centrifugation) x 100.
Time Frame	5-7 days and 35-42 days following blood donation

Outcome Measure Data

Analysis Population Description

[Not Specified]

Arm/Group Title	Biotin-Labeled Red Blood Cells Infusion
Arm/Group Description	Each participant will receive 2 transfusions of biotin labeled red blood cells. Biotin-Labeled Red Blood Cells: We will collect 500 mL of blood. The blood will be processed and split into two bags and labeled with a naturally occurring vitamin, biotin. The blood will then be re-infused back into the same participant at 2 time points (5-7 days and 35-42 days after storage).
Measure Participants	4
5-7 days	0.27 (0.09)
35-42 days	0.21 (0.14)

3. Secondary Outcome

Title	The Percentage of Red Blood Cell Osmotic Hemolysis
Description	This will be quantified by the evaluation of osmotic stress assays. We will define the associations between the primary endpoints and the quantification of osmotic hemolysis. The percent osmotic hemolysis is determined by this equation: (supernatant cell-free hemoglobin of pink test-treated RBCs / total amount of hemoglobin) x 100.
Time Frame	5-7 days and 35-42 days following blood donation

Outcome Measure Data

Analysis Population Description

[Not Specified]

Arm/Group Title	Biotin-Labeled Red Blood Cells Infusion
Arm/Group Description	Each participant will receive 2 transfusions of biotin labeled red blood cells. Biotin-Labeled Red Blood Cells: We will collect 500 mL of blood. The blood will be processed and split into two bags and labeled with a naturally occurring vitamin, biotin. The blood will then be re-infused back into the same participant at 2 time points (5-7 days and 35-42 days after storage).
Measure Participants	4
5-7 days	7.4 (4.66)
35-42 days	7.9 (6.86)

4. Secondary Outcome

Title	The Percentage of Red Blood Cell Oxidative Hemolysis
Description	This will be quantified by the evaluation of oxidative stress assays. We will define the associations between the primary endpoints and the quantification of oxidative hemolysis. The percent osmotic hemolysis is determined by this equation: (supernatant cell-free hemoglobin of 2,2'-azobis-2-methyl-propanimidamide, dihydrochloride treated RBCs - supernatant cell-free hemoglobin of untreated red blood cells / total amount of hemoglobin) x 100.
Time Frame	5-7 days and 35-42 days following blood donation

Outcome Measure Data

Analysis Population Description

[Not Specified]

Arm/Group Title	Biotin-Labeled Red Blood Cells Infusion
Arm/Group Description	Each participant will receive 2 transfusions of biotin labeled red blood cells. Biotin-Labeled Red Blood Cells: We will collect 500 mL of blood. The blood will be processed and split into two bags and labeled with a naturally occurring vitamin, biotin. The blood will then be re-infused back into the same participant at 2 time points (5-7 days and 35-42 days after storage).
Measure Participants	4
5-7 days	39.93 (12.45)
35-42 days	46.90 (21.92)

5. Post-Hoc Outcome

Title	Length of Time Biotin-labeled Red Blood Cells Can be Detected Following Infusion
Description	This will be determined using enumeration of biotinylated red blood cells obtained by flow cytometry from blood samples obtained at various time points from subjects.
Time Frame	up to 165 days

Outcome Measure Data

Analysis Population Description

The fresh stored blood sample from one participant was not collected and analyzed.

Arm/Group Title	Biotin-Labeled Red Blood Cells Infusion
Arm/Group Description	Each participant will receive 2 transfusions of biotin labeled red blood cells. Biotin-Labeled Red Blood Cells: We will collect 500 mL of blood. The blood will be processed and split into two bags and labeled with a naturally occurring vitamin, biotin. The blood will then be re-infused back into the same participant at 2 time points (5-7 days and 35-42 days after storage).
Measure Participants	4
Fresh Stored Cells	135 (12)
Stored Cells	104 (12)

Adverse Events

Time Frame	Day 125-150 post blood transfusion
Arm/Group Title	Biotin-Labeled Red Blood Cells Infusion
Arm/Group Description	Each participant will receive 2 transfusions of biotin labeled red blood cells. Biotin-Labeled Red Blood Cells: We will collect 500 mL of blood. The blood will be processed and split into two bags and labeled with a naturally occurring vitamin, biotin. The blood will then be re-infused back into the same participant at 2 time points (5-7 days and 35-42 days after storage).
All Cause Mortality
	Biotin-Labeled Red Blood Cells Infusion
	Affected / at Risk (%)	# Events
Total	0/6 (0%)
Serious Adverse Events
	Biotin-Labeled Red Blood Cells Infusion
	Affected / at Risk (%)	# Events
Total	0/6 (0%)
Other (Not Including Serious) Adverse Events
	Biotin-Labeled Red Blood Cells Infusion
	Affected / at Risk (%)	# Events
Total	1/6 (16.7%)
Blood and lymphatic system disorders
Weakly positive antibodies	1/6 (16.7%)	1

Limitations/Caveats

The clinical trial was terminated early due to challenges with enrollment and lack of study coordinator support. Early termination leading to small numbers of subjects analyzed.

More Information

Certain Agreements

All Principal Investigators ARE employed by the organization sponsoring the study.

There is NOT an agreement between Principal Investigators and the Sponsor (or its agents) that restricts the PI's rights to discuss or publish trial results after the trial is completed.