Bronchiolitis Obliterans Syndrome (BOS)NCT07018804
Mechanisms and Targeted Therapy of Airway Basal Cell Dysfunction in Bronchiolitis Obliterans Syndrome
A study testing Bronchoscopic brushing, Single - cell cloning and Cell function identification in people with Bronchiolitis Obliterans Syndrome (BOS), run by Haikou Affiliated Hospital of Central South University Xiangya School of Medicine at 1 site in China.
Trial summary
This experimental study aims to investigate the pathogenesis of bronchiolitis obliterans syndrome (BOS) and provide a basis for clinical diagnosis and treatment. The core research question is: whether there is a causal relationship between stem cell dysfunction induced by the inflammatory microenvironment and airway… Read more about this study
- Who is it for?
- Any ageHealthy volunteers can take part
- Conditions the study is for
- Bronchiolitis Obliterans Syndrome (BOS)
- What is being tested?
- Bronchoscopic brushing, Single - cell cloning and Cell function identification
- Type of treatment
- Injection or infusion, Surgery or procedure
- Could I receive a placebo?
- No, everyone receives an active treatment
- How many people
- 0 participants
- Where
- 1 siteChina
- Study dates
- 26 Jul 2026to 30 Jun 2028
- Phase
- Interventional
- Are there results?
- None posted in the registry
Key requirements
Who can take part, as listed by the sponsor. Only the study team can confirm whether you are eligible, so take this list to your doctor.
- Ages Any age
- All sexes
- Healthy volunteers accepted
- Child, Adult, Older adult
Who may be able to join
- Patients who have received allogeneic hematopoietic stem cell transplantation and are diagnosed with bronchiolitis obliterans syndrome
Who cannot join
- Bronchoscopy consultation is not suitable for patients who are not suitable for bronchoscopy.
Trial locations and contacts
1 site in 1 country.
China1 site
Haikou Affiliated Hospital of Central South University Xiangya School of Medicine
Haikou, Hainan, 570208
About this study
Official title: Analysis of the Pathogenic Mechanism of Abnormal Airway Basal Cell Function in Bronchiolitis Obliterans Syndrome and Research on Targeted Treatment Strategies
This experimental study aims to investigate the pathogenesis of bronchiolitis obliterans syndrome (BOS) and provide a basis for clinical diagnosis and treatment. The core research question is: whether there is a causal relationship between stem cell dysfunction induced by the inflammatory microenvironment and airway injury repair during the pathological process of BOS? Researchers will collect alveolar lavage fluid specimens from participants and healthy individuals to isolate distal small airway stem cells for subsequent scientific research and comparative analysis, thereby revealing the pathological mechanisms of BOS, exploring precise intervention targets, and developing innovative therapeutic strategies to improve patient prognosis, long-term survival rates, and quality of life.
Read the detailed description
The purpose of this experimental study is to explore the role of airway basal cells in the development of bronchiolitis obliterans syndrome (BOS) and develop potential therapeutic strategies. The main question it aims to answer is: How do the functional abnormalities of airway basal cells (BCs) affect the progression of BOS and what are the underlying molecular mechanisms? Researchers will collect epithelial mucosal tissues from diseased and relatively healthy lung regions of BOS patients through bronchoscopic brushing, and simultaneously gather alveolar lavage fluid specimens when possible. Specimens from healthy volunteers with no obvious airway abnormalities will be used as controls. After that, BCs will be isolated from these specimens. The isolated BCs will be cultured and their molecular characteristics will be identified using immunofluorescence staining to detect the expression of BCs markers such as p63 and Krt5.
Subsequently, single - cell clone libraries will be established by flow cytometry cell fluorescence sorting (FACS) technology. Multiple aspects of the cells' functions will be evaluated, including self - renewal ability by detecting the expression of the proliferation marker Ki67 through immunofluorescence and CCK8 assay, and differentiation ability by analyzing the cell types and proportions in the differentiation structures of in vitro air - liquid interface (ALI) culture for 21 days and in vivo differentiation in severe combined immunodeficiency (NSG) mice for 28 days using Real - time PCR and immunofluorescence techniques.
In addition, multi - omics sequencing technologies, such as RNA - seq, ATAC - seq, and CUT\&Tag, will be employed to explore the molecular mechanisms of BCs' functional abnormalities. Stable interference cell lines will be established using CRISPR - Cas9 technology to verify the functions of potential target genes. A ferret BOS model will be constructed, and BCs transplantation experiments will be carried out in ferrets. By observing and analyzing the survival rate, body weight, CT images, and lung tissue pathology of ferrets, the preventive and therapeutic effects of BCs on BOS will be evaluated.
By clarifying the role of BCs in BOS, this study aims to reveal the underlying pathological mechanisms, explore potential intervention targets, and develop novel treatment approaches. These efforts are expected to improve the prognosis of BOS patients, reduce the incidence and mortality rates, and enhance the overall quality of life for those affected by this life - threatening respiratory disorder.
- Conditions
- Bronchiolitis Obliterans Syndrome (BOS)
- Being tested
- Bronchoscopic brushing, Single - cell cloning, Cell function identification, Identification of local microenvironmental changes, Gene editing
- Keywords
- Bronchiolitis Obliterans Syndrome, Airway epithelium, Airway basal cells, Inflammatory response
- Medical subject headings
- Bronchiolitis Obliterans Syndrome
What participants receive
What participants receive. In a randomised study each person is assigned to one group by chance.
Bronchoscopic brushingProcedure or surgery
The collected samples are digested with tissue collagenase for the culture of airway basal cells (BCs).
Single - cell cloningGenetic
After primary BCs are expanded in the P1 passage, single-cell cloning libraries are established by planting them into 384-well cell culture plates in a single-cell per well format using a flow chamber cell sorter. Ten samples are selected from the expandable clones for the identification of differentiation and proliferation capabilities, while the remaining clones are cryopreserved.
Cell function identificationGenetic
For single-cell samples of each patient, in vitro expansion culture is performed to observe the morphology of cells at each passage and calculate the clonogenic rate. Immunofluorescence technique is used to detect the expression of the cell proliferation marker Ki67, and the proliferation capacity is evaluated by combining with the CCK8 assay. Cells at passages P3-P5 are seeded on the permeable membrane of cell culture inserts at a density of \(10^6\) cells/cm², and after 21 days of culture, the differentiated structures are collected. The expression of ciliated cell marker Ace-Tubulin and goblet cell marker MUC5AC is detected to assess the differentiation capacity. Meanwhile, cells are injected subcutaneously into NSG mice at \(10^6\) cells/injection site for in vivo differentiation for 28 days, and the differentiated structures are collected for pathological analysis.
Identification of local microenvironmental changesGenetic
The air-liquid interface (ALI) differentiation culture medium is collected, and cell debris is removed by high-speed centrifugation. The supernatant is then collected to extract proteins. Target proteins are purified via immunoprecipitation or affinity chromatography, followed by desalting and concentration for mass spectrometry analysis to detect inflammatory cytokines and extracellular matrix (ECM). Real-time PCR and Western blot techniques are used to measure the expression levels of epithelial markers, mesenchymal markers, and ECM in cells across all groups, thereby evaluating changes in the local microenvironment around small airways.
Gene editingGenetic
After plasmid construction, based on gene function, the CRISPR-Cas9-sgRNA (all-in-one) plasmid is transiently transfected into expanded single-cell strains via Nucleofection to knockout the target gene, or the CRISPR-dCas9 fusion-sgRNA plasmid is transiently transfected to activate or inhibit the target gene. After 3-5 days of culture, viable cells are sorted by FACS, and single cells are seeded into 96-well plates for two additional passages of expansion. Samples are collected for Sanger sequencing to screen successfully constructed cell lines.
Prevention of ferret airway basal cell transplantationProcedure or surgery
Before surgery, basal cells (BCs) of recipient ferrets are collected by bronchoscopic brushing, followed by in vitro culture and identification. Prior to the onset of BOS, the cells are injected into recipient ferrets via bronchoscopy. Outcomes including ferret survival rate, body weight, CT imaging, and lung tissue pathology are collected to evaluate the preventive effect of BCs on BOS.
Transplantation treatment of ferret airway basal cellsProcedure or surgery
Stable cell lines with target gene knockdown are established in ferret basal cells (BCs). After ferrets develop bronchiolitis obliterans syndrome (BOS), the constructed cell lines are transplanted into recipient ferrets to validate the therapeutic effect of gene-corrected BCs on the disease.
| Study group | Role | What they receive |
|---|---|---|
| Patient group | Experimental | Epithelial mucosal tissues from the diseased lung regions and relatively healthy lung lobes of BOS patients following hematopoietic stem cell transplantation (HSCT) will be collected. Alveolar lavage fluid specimens will be obtained when available. The intervention consists of digesting the collected samples with tissue collagenase for the culture of airway basal cells (BCs). Receives: Procedure: Bronchoscopic brushing |
| Healthy control group | Experimental | Healthy volunteers with no obvious airway abnormalities will be selected to collect samples from the same anatomical sites. No disease-related interventions will be performed, serving as a comparative control to highlight the abnormal conditions of basal cells (BCs) in the patient group. Receives: Procedure: Bronchoscopic brushing |
| Single - cell clone group | Experimental | Not described Receives: Genetic: Single - cell cloning |
| Cell Function Identification Group | Experimental | Not described Receives: Genetic: Cell function identification |
| Local microenvironment change identification group | Experimental | Not described Receives: Genetic: Identification of local microenvironmental changes |
| Gene Editing Group | Experimental | Not described Receives: Genetic: Gene editing |
| Ferret airway basal cell transplantation group (prevention group) | Experimental | Not described Receives: Procedure: Prevention of ferret airway basal cell transplantation |
| Ferret airway basal cell transplantation group (treatment group) | Experimental | Not described Receives: Procedure: Transplantation treatment of ferret airway basal cells |
What is being measured
The primary outcome is the main question the study is designed to answer.
Ki67 Expression Level in Airway Basal Cells
The percentage of Ki67-positive airway basal cells assessed by immunohistochemistry. Ki67 is a nuclear protein associated with cellular proliferation. Higher Ki67 expression indicates greater proliferative activity. Comparison will be made between patients with bronchiolitis obliterans syndrome and healthy controls to evaluate abnormal self-renewal function of airway basal cells in disease states.
Measured: From enrollment to 32 weeks
Cell Proliferative Activity Assessed by CCK8 Assay
Optical density (OD) values measured by the CCK8 (Cell Counting Kit-8) assay in cultured airway basal cells. The CCK8 assay measures cell viability and proliferation based on the reduction of WST-8 by dehydrogenases in living cells. Higher OD values indicate greater cell proliferation. Comparison will be made between patients with bronchiolitis obliterans syndrome and healthy controls.
Measured: From enrollment to 32 weeks
Expression of Ciliated Cell Marker Ace-Tubulin in Air-Liquid Interface (ALI) Differentiation Culture
Immunofluorescence staining intensity of Ace-Tubulin (acetylated α-tubulin), a marker for ciliated cells, in airway basal cells cultured under air-liquid interface (ALI) conditions for 21 days. The staining intensity reflects the degree of ciliated cell differentiation from airway basal cells.
Measured: From enrollment to 32 weeks
Expression of Goblet Cell Marker MUC5AC in Air-Liquid Interface (ALI) Differentiation Culture
Immunofluorescence staining intensity of MUC5AC, a marker for goblet cells, in airway basal cells cultured under air-liquid interface (ALI) conditions for 21 days. The staining intensity reflects the degree of goblet cell differentiation from airway basal cells.
Measured: From enrollment to 32 weeks
Expression of Ciliated Cell Marker Ace-Tubulin in NSG Mouse Xenograft Model
Immunofluorescence staining intensity of Ace-Tubulin (acetylated α-tubulin), a marker for ciliated cells, in airway basal cells differentiated in vivo in severe combined immunodeficiency (NSG) mice for 28 days.
Measured: From enrollment to 32 weeks
Expression of Goblet Cell Marker MUC5AC in NSG Mouse Xenograft Model
Immunofluorescence staining intensity of MUC5AC, a marker for goblet cells, in airway basal cells differentiated in vivo in severe combined immunodeficiency (NSG) mice for 28 days.
Measured: From enrollment to 32 weeks
Survival Rate of Ferrets After Airway Basal Cell Transplantation
Percentage of ferrets surviving at the end of the 32-week observation period following airway basal cell transplantation. Survival is recorded daily. This indicator evaluates the impact of airway basal cell transplantation on disease progression in the bronchiolitis obliterans syndrome ferret model.
Measured: From enrollment to 32 weeks
Body Weight Changes in Ferrets After Airway Basal Cell Transplantation
Body weight of ferrets measured at baseline and at regular intervals (weekly) over the 32-week observation period following airway basal cell transplantation. Weight change is calculated as the percentage change from baseline. This indicator reflects the overall health status of ferrets and assists in evaluating the effect of airway basal cell transplantation.
Measured: From enrollment to 32 weeks
Secondary outcomes (1)
Expression of Airway Basal Cell Markers p63 and Krt5 by Immunofluorescence
Immunofluorescence staining of transformation protein 63 (p63) and keratin 5 (Krt5), two specific markers for airway basal cells. The percentage of p63-positive and Krt5-positive cells among total cultured cells will be assessed. Positive expression of both markers confirms the identity of cultured cells as airway basal cells, ensuring cell source quality for subsequent experiments.
Measured: From enrollment to 32 weeks
Timeline
From the start of the study to its completion; the mark is today. Estimated dates are the sponsor’s plan.
- First posted12 Jun 2025
- Study start26 Jul 2026 (estimated)
- Primary completion30 Jun 2027 (estimated)
- Study completion30 Jun 2028 (estimated)
Why it stopped: Study was withdrawn prior to enrollment due to dissolution of collaborative partnership. The protocol may be reactivated in the future under new collaboration.
Resources
- Official record on ClinicalTrials.govThe registry entry this page is built from, with its full change history.
- Other trials for Bronchiolitis Obliterans Syndrome (BOS)Studies of the same condition, open and under way.
- What the trial phases meanHow a treatment moves from first tests in people to approval.
More details
For doctors and researchers: how the study is designed, who runs it, and its registry IDs.
Study design
- Study type
- Interventional
- Phase
- Phase N/A
- Primary purpose
- Supportive care
- Allocation
- Non-randomised
- Intervention model
- Parallel groups
- Model description
- In this experiment, bronchoscopic brushing was used to collect epithelial mucosal tissues from the diseased lung area and the relatively healthy lung lobe area of the patients. When conditions permit, alveolar lavage fluid specimens of the patients were collected simultaneously to isolate distal small airway stem cells. Allelic samples from healthy volunteers with no obvious airway abnormalities were used as parallel controls. The collected samples were digested with tissue collagenase and then used for the culture of airway BCS.
- Masking
- None (open label)
- Enrolment
- 0 participants (actual)
Sponsor and investigators
- Lead sponsor
- Haikou Affiliated Hospital of Central South University Xiangya School of Medicine
- Sponsor type
- University, hospital or other
- Responsible party
- Sponsor
- Investigators
- Xiaoyang Yang, DM, Principal investigator, Department of Hematology, Haikou People's Hospital
IDs, dates and publications
- ClinicalTrials.gov ID
- NCT07018804
- Sponsor's protocol ID
- SC20250160
- Status last verified
- Jul 2026
- First submitted
- 17 May 2025
- Last updated
- 29 Jul 2026
- Results posted
- No
- Data monitoring committee
- Yes
- FDA-regulated drug
- No
- FDA-regulated device
- No
- Individual data sharing
- No: The IPD encompasses highly sensitive and confidential data that was collected through significant investment of our resources, both in terms of time and funding. This data is integral to our ongoing research initiatives, which are at a crucial stage of development. Premature sharing could disrupt our research timelines and strategic plans. Additionally, we have not yet established comprehensive safeguards to ensure that the data will be used appropriately by external researchers. Without proper protocols in place, there is a risk of misuse or misinterpretation of the data, which could lead to inaccurate research outcomes and potential reputational damage to our institution. For these reasons, we have decided not to share the IPD at this time.
Study at a glance
From the registry record, last updated by the sponsor on 29 Jul 2026. Recruitment status is as the sponsor reported it: confirm with the study team.
- Official title
- Analysis of the Pathogenic Mechanism of Abnormal Airway Basal Cell Function in Bronchiolitis Obliterans Syndrome and Research on Targeted Treatment Strategies
- ClinicalTrials.gov ID
- NCT07018804
- Acronym / sponsor ID
- SC20250160
- Conditions
- Bronchiolitis Obliterans Syndrome (BOS)
- Keywords
- Bronchiolitis Obliterans Syndrome, Airway epithelium, Airway basal cells, Inflammatory response
- Study type
- Interventional
- Recruitment status
- WithdrawnReason stopped: Study was withdrawn prior to enrollment due to dissolution of collaborative partnership. The protocol may be reactivated in the future under new collaboration.
- Enrolment
- 0 participants (actual)
- Lead sponsor
- Haikou Affiliated Hospital of Central South University Xiangya School of MedicineUniversity, hospital or other
- Study start
- 26 Jul 2026 (estimated)
- Primary completion
- 30 Jun 2027 (estimated)
- Study completion
- 30 Jun 2028 (estimated)
- First posted
- 12 Jun 2025 (actual)
- Results
- Not posted in the source record
Clinical summary
The research question as the sponsor registered it. This describes what the study plans to test, not what it found.
This experimental study aims to investigate the pathogenesis of bronchiolitis obliterans syndrome (BOS) and provide a basis for clinical diagnosis and treatment. The core research question is: whether there is a causal relationship between stem cell dysfunction induced by the inflammatory microenvironment and airway injury repair during the pathological process of BOS? Researchers will collect alveolar lavage fluid specimens from participants and healthy individuals to isolate distal small airway stem cells for subsequent scientific research and comparative analysis, thereby revealing the pathological mechanisms of BOS, exploring precise intervention targets, and developing innovative therapeutic strategies to improve patient prognosis, long-term survival rates, and quality of life.
Detailed description
The purpose of this experimental study is to explore the role of airway basal cells in the development of bronchiolitis obliterans syndrome (BOS) and develop potential therapeutic strategies. The main question it aims to answer is: How do the functional abnormalities of airway basal cells (BCs) affect the progression of BOS and what are the underlying molecular mechanisms? Researchers will collect epithelial mucosal tissues from diseased and relatively healthy lung regions of BOS patients through bronchoscopic brushing, and simultaneously gather alveolar lavage fluid specimens when possible. Specimens from healthy volunteers with no obvious airway abnormalities will be used as controls. After that, BCs will be isolated from these specimens. The isolated BCs will be cultured and their molecular characteristics will be identified using immunofluorescence staining to detect the expression of BCs markers such as p63 and Krt5.
Subsequently, single - cell clone libraries will be established by flow cytometry cell fluorescence sorting (FACS) technology. Multiple aspects of the cells' functions will be evaluated, including self - renewal ability by detecting the expression of the proliferation marker Ki67 through immunofluorescence and CCK8 assay, and differentiation ability by analyzing the cell types and proportions in the differentiation structures of in vitro air - liquid interface (ALI) culture for 21 days and in vivo differentiation in severe combined immunodeficiency (NSG) mice for 28 days using Real - time PCR and immunofluorescence techniques.
In addition, multi - omics sequencing technologies, such as RNA - seq, ATAC - seq, and CUT\&Tag, will be employed to explore the molecular mechanisms of BCs' functional abnormalities. Stable interference cell lines will be established using CRISPR - Cas9 technology to verify the functions of potential target genes. A ferret BOS model will be constructed, and BCs transplantation experiments will be carried out in ferrets. By observing and analyzing the survival rate, body weight, CT images, and lung tissue pathology of ferrets, the preventive and therapeutic effects of BCs on BOS will be evaluated.
By clarifying the role of BCs in BOS, this study aims to reveal the underlying pathological mechanisms, explore potential intervention targets, and develop novel treatment approaches. These efforts are expected to improve the prognosis of BOS patients, reduce the incidence and mortality rates, and enhance the overall quality of life for those affected by this life - threatening respiratory disorder.
Study design
- Primary purpose
- Supportive care
- Allocation
- Non-randomised
- Intervention model
- Parallel groups
- Masking
- None (open label)
- Number of arms
- 8
- Model description
- In this experiment, bronchoscopic brushing was used to collect epithelial mucosal tissues from the diseased lung area and the relatively healthy lung lobe area of the patients. When conditions permit, alveolar lavage fluid specimens of the patients were collected simultaneously to isolate distal small airway stem cells. Allelic samples from healthy volunteers with no obvious airway abnormalities were used as parallel controls. The collected samples were digested with tissue collagenase and then used for the culture of airway BCS.
| Arm | Type | Receives |
|---|---|---|
| Patient group | Experimental | Procedure: Bronchoscopic brushingEpithelial mucosal tissues from the diseased lung regions and relatively healthy lung lobes of BOS patients following hematopoietic stem cell transplantation (HSCT) will be collected. Alveolar lavage fluid specimens will be obtained when available. The intervention consists of digesting the collected samples with tissue collagenase for the culture of airway basal cells (BCs). |
| Healthy control group | Experimental | Procedure: Bronchoscopic brushingHealthy volunteers with no obvious airway abnormalities will be selected to collect samples from the same anatomical sites. No disease-related interventions will be performed, serving as a comparative control to highlight the abnormal conditions of basal cells (BCs) in the patient group. |
| Single - cell clone group | Experimental | Genetic: Single - cell cloning |
| Cell Function Identification Group | Experimental | Genetic: Cell function identification |
| Local microenvironment change identification group | Experimental | Genetic: Identification of local microenvironmental changes |
| Gene Editing Group | Experimental | Genetic: Gene editing |
| Ferret airway basal cell transplantation group (prevention group) | Experimental | Procedure: Prevention of ferret airway basal cell transplantation |
| Ferret airway basal cell transplantation group (treatment group) | Experimental | Procedure: Transplantation treatment of ferret airway basal cells |
Who may qualify
- Sex
- All
- Healthy volunteers
- Accepted
- Age groups
- Child, Adult, Older adult
Key inclusion criteria (1 listed)
- Patients who have received allogeneic hematopoietic stem cell transplantation and are diagnosed with bronchiolitis obliterans syndrome
Key exclusion criteria (1 listed)
- Bronchoscopy consultation is not suitable for patients who are not suitable for bronchoscopy.
Eligibility is decided by the study team after screening; this overview does not establish that a patient qualifies. Full inclusion and exclusion criteria.
Interventions
| Intervention | Type | Arms |
|---|---|---|
| Bronchoscopic brushingThe collected samples are digested with tissue collagenase for the culture of airway basal cells (BCs). | Procedure or surgery | Healthy control group, Patient group |
| Single - cell cloningAfter primary BCs are expanded in the P1 passage, single-cell cloning libraries are established by planting them into 384-well cell culture plates in a single-cell per well format using a flow chamber cell sorter. Ten samples are selected from the expandable clones for the identification of differentiation and proliferation capabilities, while the remaining clones are cryopreserved. | Genetic | Single - cell clone group |
| Cell function identificationFor single-cell samples of each patient, in vitro expansion culture is performed to observe the morphology of cells at each passage and calculate the clonogenic rate. Immunofluorescence technique is used to detect the expression of the cell proliferation marker Ki67, and the proliferation capacity is evaluated by combining with the CCK8 assay. Cells at passages P3-P5 are seeded on the permeable membrane of cell culture inserts at a density of \(10^6\) cells/cm², and after 21 days of culture, the differentiated structures are collected. The expression of ciliated cell marker Ace-Tubulin and goblet cell marker MUC5AC is detected to assess the differentiation capacity. Meanwhile, cells are injected subcutaneously into NSG mice at \(10^6\) cells/injection site for in vivo differentiation for 28 days, and the differentiated structures are collected for pathological analysis. | Genetic | Cell Function Identification Group |
| Identification of local microenvironmental changesThe air-liquid interface (ALI) differentiation culture medium is collected, and cell debris is removed by high-speed centrifugation. The supernatant is then collected to extract proteins. Target proteins are purified via immunoprecipitation or affinity chromatography, followed by desalting and concentration for mass spectrometry analysis to detect inflammatory cytokines and extracellular matrix (ECM). Real-time PCR and Western blot techniques are used to measure the expression levels of epithelial markers, mesenchymal markers, and ECM in cells across all groups, thereby evaluating changes in the local microenvironment around small airways. | Genetic | Local microenvironment change identification group |
| Gene editingAfter plasmid construction, based on gene function, the CRISPR-Cas9-sgRNA (all-in-one) plasmid is transiently transfected into expanded single-cell strains via Nucleofection to knockout the target gene, or the CRISPR-dCas9 fusion-sgRNA plasmid is transiently transfected to activate or inhibit the target gene. After 3-5 days of culture, viable cells are sorted by FACS, and single cells are seeded into 96-well plates for two additional passages of expansion. Samples are collected for Sanger sequencing to screen successfully constructed cell lines. | Genetic | Gene Editing Group |
| Prevention of ferret airway basal cell transplantationBefore surgery, basal cells (BCs) of recipient ferrets are collected by bronchoscopic brushing, followed by in vitro culture and identification. Prior to the onset of BOS, the cells are injected into recipient ferrets via bronchoscopy. Outcomes including ferret survival rate, body weight, CT imaging, and lung tissue pathology are collected to evaluate the preventive effect of BCs on BOS. | Procedure or surgery | Ferret airway basal cell transplantation group (prevention group) |
| Transplantation treatment of ferret airway basal cellsStable cell lines with target gene knockdown are established in ferret basal cells (BCs). After ferrets develop bronchiolitis obliterans syndrome (BOS), the constructed cell lines are transplanted into recipient ferrets to validate the therapeutic effect of gene-corrected BCs on the disease. | Procedure or surgery | Ferret airway basal cell transplantation group (treatment group) |
Primary outcome
Ki67 Expression Level in Airway Basal Cells
Time frame: From enrollment to 32 weeks
The percentage of Ki67-positive airway basal cells assessed by immunohistochemistry. Ki67 is a nuclear protein associated with cellular proliferation. Higher Ki67 expression indicates greater proliferative activity. Comparison will be made between patients with bronchiolitis obliterans syndrome and healthy controls to evaluate abnormal self-renewal function of airway basal cells in disease states.
Cell Proliferative Activity Assessed by CCK8 Assay
Time frame: From enrollment to 32 weeks
Optical density (OD) values measured by the CCK8 (Cell Counting Kit-8) assay in cultured airway basal cells. The CCK8 assay measures cell viability and proliferation based on the reduction of WST-8 by dehydrogenases in living cells. Higher OD values indicate greater cell proliferation. Comparison will be made between patients with bronchiolitis obliterans syndrome and healthy controls.
Expression of Ciliated Cell Marker Ace-Tubulin in Air-Liquid Interface (ALI) Differentiation Culture
Time frame: From enrollment to 32 weeks
Immunofluorescence staining intensity of Ace-Tubulin (acetylated α-tubulin), a marker for ciliated cells, in airway basal cells cultured under air-liquid interface (ALI) conditions for 21 days. The staining intensity reflects the degree of ciliated cell differentiation from airway basal cells.
Expression of Goblet Cell Marker MUC5AC in Air-Liquid Interface (ALI) Differentiation Culture
Time frame: From enrollment to 32 weeks
Immunofluorescence staining intensity of MUC5AC, a marker for goblet cells, in airway basal cells cultured under air-liquid interface (ALI) conditions for 21 days. The staining intensity reflects the degree of goblet cell differentiation from airway basal cells.
Expression of Ciliated Cell Marker Ace-Tubulin in NSG Mouse Xenograft Model
Time frame: From enrollment to 32 weeks
Immunofluorescence staining intensity of Ace-Tubulin (acetylated α-tubulin), a marker for ciliated cells, in airway basal cells differentiated in vivo in severe combined immunodeficiency (NSG) mice for 28 days.
Expression of Goblet Cell Marker MUC5AC in NSG Mouse Xenograft Model
Time frame: From enrollment to 32 weeks
Immunofluorescence staining intensity of MUC5AC, a marker for goblet cells, in airway basal cells differentiated in vivo in severe combined immunodeficiency (NSG) mice for 28 days.
Survival Rate of Ferrets After Airway Basal Cell Transplantation
Time frame: From enrollment to 32 weeks
Percentage of ferrets surviving at the end of the 32-week observation period following airway basal cell transplantation. Survival is recorded daily. This indicator evaluates the impact of airway basal cell transplantation on disease progression in the bronchiolitis obliterans syndrome ferret model.
Body Weight Changes in Ferrets After Airway Basal Cell Transplantation
Time frame: From enrollment to 32 weeks
Body weight of ferrets measured at baseline and at regular intervals (weekly) over the 32-week observation period following airway basal cell transplantation. Weight change is calculated as the percentage change from baseline. This indicator reflects the overall health status of ferrets and assists in evaluating the effect of airway basal cell transplantation.
1 secondary outcome
Expression of Airway Basal Cell Markers p63 and Krt5 by Immunofluorescence
Time frame: From enrollment to 32 weeks
Results and safety
The study was withdrawn before enrolling participants, so there are no results.
Research links
- Official record on ClinicalTrials.govLast updated by the sponsor 29 Jul 2026.
- Individual participant dataSharing planned: No. The IPD encompasses highly sensitive and confidential data that was collected through significant investment of our resources, both in terms of time and funding. This data is integral to our ongoing research initiatives, which are at a crucial stage of development. Premature sharing could disrupt our research timelines and strategic plans. Additionally, we have not yet established comprehensive safeguards to ensure that the data will be used appropriately by external researchers. Without proper protocols in place, there is a risk of misuse or misinterpretation of the data, which could lead to inaccurate research outcomes and potential reputational damage to our institution. For these reasons, we have decided not to share the IPD at this time.
Related trials for Bronchiolitis Obliterans Syndrome (BOS)
RecruitingPhase 2
Efficacy of an Anti-TSLP Monoclonal Antibody in the Management of Chronic Bronchial Disease Induced by Bronchiolitis Obliterans Syndrome in Allogeneic Hematopoietic Stem Cells Transplantation Recipients
Bronchiolitis Obliterans Syndrome (BOS), HSCT
RecruitingPhase 3
Study to Evaluate ARINA-1 in the Prevention of Bronchiolitis Obliterans Progression in Participants With Bilateral Lung Transplant
Pre-Bronchiolitis Obliterans Syndrome
Recruiting
Forced Oscillatory Technique for Bronchiolitis Obliterans Syndrome After Allogeneic Bone Marrow Transplantation
Bronchiolitis Obliterable Syndrome
Active, not recruitingObservational study
The Longitudinal Impact of Respiratory Viruses on Bronchiolitis Obliterans Syndrome After Allogeneic Hematopoietic Cell Transplantation (The RV-BOS Study)
Bronchiolitis Obliterans Syndrome, Hematopoietic and Lymphoid Cell Neoplasm
RecruitingObservational study
Plasma and Radiologic Biomarkers of Response to ECP in Lung Transplant Recipients With CLAD
Lung Transplant Failure and Rejection, CLAD, Bronchiolitis Obliterans
Referral and contact
AzTrials does not enrol participants or take referrals. Contact the study team or a site listed below; they decide who can take part. This study is not recruiting at the moment.
Xiaoyang Yang, DM principal investigator
Department of Hematology, Haikou People's Hospital
1 site in 1 country: see every site and its status.
Questions about this trial
What is this clinical trial studying?
A study testing Bronchoscopic brushing, Single - cell cloning and Cell function identification in people with Bronchiolitis Obliterans Syndrome (BOS), run by Haikou Affiliated Hospital of Central South University Xiangya School of Medicine at 1 site in China.
The full registered description is in the About this study section and the official record.
Who may be eligible to take part?
The sponsor lists 1 inclusion and 1 exclusion criteria. Eligibility is decided by the study team after screening, so read the full criteria and contact a listed site.
Is this clinical trial currently recruiting?
The record says: withdrawn, as of the sponsor's last update on 29 Jul 2026. Status can change between updates, so confirm with the study contact before making plans.
Where is the study taking place?
The record lists 1 site in China. Whether a site is taking part right now should be confirmed with that site. See every site.
What are the possible benefits and risks?
They depend on the study and the treatment, and no study can promise a benefit. The study team explains the known risks during informed consent, before anyone joins; the record itself does not list them.
Are results available for this study?
The study was withdrawn before enrolling participants, so there are no results.
How can a doctor refer a patient or contact the research team?
The record lists no contact. The official record names the sponsor, who can be asked directly. AzTrials does not enrol participants or take referrals.
What does the primary outcome measure?
Ki67 Expression Level in Airway Basal Cells (time frame: From enrollment to 32 weeks). The study lists 8 primary outcomes in all. It is what the study was designed to measure, not a finding.
Where can I read the original study information?
Does a listing mean the treatment is approved or proven to work?
No. A listing means the study is registered. It does not show that the treatment is safe, works, or is approved by a regulator.
How often is this information updated?
AzTrials copies changed records from ClinicalTrials.gov every night and reloads the whole registry every week. The sponsor last updated this record on 29 Jul 2026.