The Mechanism of Enhancing the Anti-tumor Effects of CAR-T on PC by Gut Microbiota Regulation

December 16, 2019 updated by: First Affiliated Hospital of Harbin Medical University

Study on the Mechanism of Enhancing the Anti-tumor Effects of Human Chimeric Antigen Receptors T Cells on Pancreatic Cancer by Gut Microbiota Regulation

Pancreatic cancer (PC) is one of the deadliest diseases of human digestive malignancies. Despite the recent advances in surgery and chemotherapy, the 5-year survival rate of PC continues to be less than 10%. As a promising tumor therapy,Chimeric antigen receptor T cell (CAR-T), however, performed poorly in PC treatment and need to be further updated. In our study, on the basis of our previous research, we use anti-MSLN CAR-T as effector cell and explore the different effects and mechanism of gut microbiota (PC or healthy control) on anti-MSLN CAR-T treatment. Firstly, we detect the differences of gut microbiota and T cell cholesterol metabolism in PC and healthy control by means of 16S-rRNA,PCR, western blot and ELISA; explore the different effects of gut microbiota on the subtype of T cells; and analyze the relationships between intestinal flora composition and T cell cholesterol metabolism or subtype changes by means of Spearman's correlation. Secondly, we also explore the different effects of gut microbiota on the proliferation, migration, subtype, inflammatory cytokines expression and anti-tumor effector function of anti-MSLN CAR-T cells by means of flow cytometry and cytotoxicity assay. Thirdly, we discuss the different expression of cholesterol esterification enzyme 1 (ACAT-1) and other core genes of cholesterol metabolism in anti-MSLN CAR-T. Lastly, we evaluate the effects of different gut microbiota on the treatment of PC by anti-MSLN CAR-T cells in NSG mouse model of subcutaneous PC transplantation and liver metastasis. Through the above experiments, a new theoretical basis is provided in which gut microbiota regulates the subtype and anti-tumor function of anti-MSLN CAR-T by ACAT-1 expression. Furthermore, our findings, which demonstrate the relationship of gut microbiota and CAR-T cell, may be translatable for the treatment of other solid tumors like PC.

Study Overview

Status

Status

Indicates the current recruitment status or the expanded access status.
Unknown

Conditions

Conditions

The disease, disorder, syndrome, illness, or injury that is being studied. On ClinicalTrials.gov, conditions may also include other health-related issues, such as lifespan, quality of life, and health risks.

Study Type

Study Type

Describes the nature of a clinical study. Study types include interventional studies (also called clinical trials), observational studies (including patient registries), and expanded access.
Observational

Enrollment (Anticipated)

Enrollment

The number of participants in a clinical study. The "anticipated" enrollment is the target number of participants that the researchers need for the study.
80

Contacts and Locations

This section provides the contact details for those conducting the study, and information on where this study is being conducted.

Study Contact

Study Contact

The name and contact information for the person who can answer enrollment questions for a clinical study. Each location where the study is being conducted may also have a specific contact, who may be better able to answer those questions.

Study Contact Backup

Study Locations

  • China
    • Heilongjiang
      • Harbin, Heilongjiang, China, 150081
        • Recruiting
        • The First Affiliated Hospital of Harbin Medical University
        • Contact:

Participation Criteria

Researchers look for people who fit a certain description, called eligibility criteria. Some examples of these criteria are a person's general health condition or prior treatments.

Eligibility Criteria

Eligibility Criteria

The key requirements that people who want to participate in a clinical study must meet or the characteristics they must have. Eligibility criteria consist of both inclusion criteria (which are required for a person to participate in the study) and exclusion criteria (which prevent a person from participating). Types of eligibility criteria include whether a study accepts healthy volunteers, has age or age group requirements, or is limited by sex.

Ages Eligible for Study

30 years to 75 years (Adult, Older Adult)

Accepts Healthy Volunteers

Yes

Genders Eligible for Study

All

Sampling Method

Non-Probability Sample

Study Population

Pancreatic cancer patients and Healthy person.

Description

1. Inclusion criteria and exclusion criteria for the experimental group of this project

Inclusion Criteria:

  1. the patients who are confirmed by imaging, pathology and body fluid biopsy to have metastasized pancreatic cancer and can not be cured by operation; or the patients who recover well one month after operation but still have residual focus, recurrence or metastasis.
  2. age ≥ 30 and ≤ 75
  3. the estimated life span is more than 1 month.
  4. Karnofsky score ≥ 60; ECoG ≤ 2
  5. the function of important organs: Echocardiography indicated that the ejection fraction of heart was ≥ 50%; ECG showed no obvious abnormality; creatinine clearance rate calculated by Cockcroft Gault formula was ≥ 40ml / min; ALT and AST were ≤ 3 times of normal value; total bilirubin was ≤ 2.0mg/dl; coagulation function: Pt and appt were < 2 times of normal value; arterial oxygen saturation (SpO2) > 92%;Blood routine test: Hgb ≥ 80g / L, ANC ≥ 1 × 109 / L, PLT ≥ 50 × 109 / L.
  6. sign the informed consent.

Exclusion Criteria:

  1. patients who used immunosuppressive drugs, hormones, antibiotics and probiotics one month before admission.
  2. serious active infection.
  3. HIV positive, active hepatitis infection.
  4. previous history of other malignant tumors. Excluding: Patients with cured skin basal or squamous cell carcinoma and cervical carcinoma in situ at any time before the study; patients with other tumors not listed above but cured by operation only without other further treatment measures and disease-free survival period ≥ 5 years can be included in the study.

(6) patients participating in other clinical trials at the same time. (7) in the opinion of the researcher, the subjects are not suitable to be selected or cannot cooperate to participate in or complete the study.

(8) patients with congenital immunodeficiency. Exclusion criteria

  1. patients who had used immunosuppressive drugs, hormones, antibiotics and probiotics in the first month before inclusion.
  2. severe active infection.
  3. human immunodeficiency virus (HIV) positive, active hepatitis infection.
  4. previous history of other malignant tumors. Excluding: patients with cured basal or squamous cell carcinoma of the skin and carcinoma in situ of the cervix at any time prior to the study; Subjects with disease-free survival ≥5 years of other tumors not listed above, which have been cured by surgery only without other further treatment measures, can be included in the study.

(6) patients who also participate in other clinical trials. (7) the investigator considers that the subject is not suitable for inclusion or unable to cooperate in or complete the study.

(8) patients with congenital immune deficiency. (9) a history of myocardial infarction and severe arrhythmia within six months. 2. Inclusion criteria and exclusion criteria for the control group of this project

  1. no history of digestive tract diseases, infectious diseases or immune diseases.
  2. no history of smoking or drinking.
  3. did not take antibiotics and other drugs and probiotics for 1 month before enrollment.

Study Plan

This section provides details of the study plan, including how the study is designed and what the study is measuring.

How is the study designed?

Design Details

Cohorts and Interventions

Group / Cohort

Group / Cohort

A group or subgroup of participants in an observational study that is assessed for biomedical or health outcomes.
Pancreatic cancer patient
(1) patients with pancreatic cancer confirmed by imaging, pathology and body fluid biopsy, or patients who recovered well one month after operation but still had residual lesions, recurrence or metastasis. ② age ≥ 30 and ≤ 75. ③ no chemotherapy, radiotherapy or targeted drugs were used before admission. ④ patients who had used immunosuppressive drugs, hormones, antibiotics and probiotics one month before admission were excluded. ⑤ patients with HIV positive and active hepatitis B or C infection were excluded. ⑥ exclude the previous history of other malignant tumors or the current combination of other malignant tumors. The patients with serious cardiopulmonary disease and liver and kidney dysfunction were excluded. ⑧ patients with obstructive jaundice were excluded.
Healthy person
(1) no history of digestive tract diseases, infectious diseases or immune diseases.(2) no history of smoking or drinking.(3) did not take antibiotics and other drugs and probiotics for 1 month before enrollment.

What is the study measuring?

Primary Outcome Measures

Primary Outcome Measures

In a clinical study's protocol, the planned outcome measure that is the most important for evaluating the effect of an intervention/treatment. Most clinical studies have one primary outcome measure, but some have more than one.
Outcome Measure
Measure Description
Time Frame
In vivo experiment: comparison of intestinal microflora and T cell cholesterol metabolism and subtypes between pancreatic cancer patients and healthy people The difference of proportion and the correlation.
Time Frame: from January 2020 to December 2020.

Fecal and peripheral blood samples were collected from patients with pancreatic cancer and healthy people respectively: (1) through Illumina miseq

High throughput sequencing technology was used to detect 16S rRNA to analyze the diversity of intestinal flora (α and β diversity), and bioinformatics methods such as random forest and lefse were used to analyze the diversity of flora composition; (2) the difference of cholesterol content in T cells was detected by ELISA; (3) the difference of ACAT-1 expression in T cells was detected by PCR and WB; (4) the difference of ACAT-1 expression in T cells was detected by flow cytometry. The proportion of CD8 +, TNF α +, Th1, Th2, Th17, Treg and other subtypes in T cells was detected by cell analyzer. Furthermore, the correlation between intestinal microflora composition and peripheral blood T cell cholesterol metabolism and subtype difference was determined by Spearman correlation analysis (statistical method).
from January 2020 to December 2020.
In vitro experiment: to verify the regulation of intestinal flora on the biological behavior of anti msln car-t in vitro and the role of ACAT-1 in it.
Time Frame: from January 2021 to December 2021.
In vitro, intestinal flora supernatant of pancreatic cancer and healthy human were co-cultured with anti-msln car-t, respectively, to verify the effect of intestinal flora on biological behavior of anti-msln car-t.To analyze how the metabolism of cholesterol in anti-msln car-t cells, with acat-1 as the key enzyme, participates in the process of regulating the biological behavior of anti-msln car-t by intestinal flora, and preliminarily uncover the relevant mechanism.
from January 2021 to December 2021.
Animal model experiment: to verify the ability of intestinal bacteria to control anti msln car-t targeted killing pancreatic cancer cells.
Time Frame: from January 2022 to December 2022.
Furthermore, the influence of differences in intestinal flora composition between pancreatic cancer and healthy human on anti-msln car-t function was verified in animal models of pancreatic cancer.
from January 2022 to December 2022.

Collaborators and Investigators

This is where you will find people and organizations involved with this study.

Sponsor

Sponsor

The organization or person who initiates the study and who has authority and control over the study.
First Affiliated Hospital of Harbin Medical University

Study record dates

These dates track the progress of study record and summary results submissions to ClinicalTrials.gov. Study records and reported results are reviewed by the National Library of Medicine (NLM) to make sure they meet specific quality control standards before being posted on the public website.

Study Major Dates

Study Start (Actual)

Study Start

The actual date on which the first participant was enrolled in a clinical study. The "anticipated" study start date is the date that the researchers think will be the study start date.
October 20, 2019

Primary Completion (Anticipated)

Primary Completion

The date on which the last participant in a clinical study was examined or received an intervention to collect final data for the primary outcome measure. Whether the clinical study ended according to the protocol or was terminated does not affect this date. For clinical studies with more than one primary outcome measure with different completion dates, this term refers to the date on which data collection is completed for all the primary outcome measures. The "anticipated" primary completion date is the date that the researchers think will be the primary completion date for the study.
December 31, 2022

Study Completion (Anticipated)

Study Completion

The date on which the last participant in a clinical study was examined or received an intervention/treatment to collect final data for the primary outcome measures, secondary outcome measures, and adverse events (that is, the last participant's last visit). The "anticipated" study completion date is the date that the researchers think will be the study completion date.
December 31, 2022

Study Registration Dates

First Submitted

First Submitted

The date on which the study sponsor or investigator first submitted a study record to ClinicalTrials.gov. There is typically a delay of a few days between the first submitted date and the record's availability on ClinicalTrials.gov (the first posted date).
December 16, 2019

First Submitted That Met QC Criteria

First Submitted That Met QC Criteria

The date on which the study sponsor or investigator first submits a study record that is consistent with National Library of Medicine (NLM) quality control (QC) review criteria. The sponsor or investigator may need to revise and submit a study record one or more times before NLM's QC review criteria are met. It is the responsibility of the sponsor or investigator to ensure that the study record is consistent with the NLM QC review criteria.
December 16, 2019

First Posted (Actual)

First Posted

The date on which the study record was first available on ClinicalTrials.gov after National Library of Medicine (NLM) quality control (QC) review has concluded. There is typically a delay of a few days between the date the study sponsor or investigator submitted the study record and the first posted date.
December 18, 2019

Study Record Updates

Last Update Posted (Actual)

Last Update Posted

The most recent date on which changes to a study record were made available on ClinicalTrials.gov. There may be a delay between when the changes were submitted to ClinicalTrials.gov by the study's sponsor or investigator (the last update submitted date) and the last update posted date.
December 18, 2019

Last Update Submitted That Met QC Criteria

Last Update Submitted That Met QC Criteria

The most recent date on which the study sponsor or investigator submitted changes to a study record that are consistent with National Library of Medicine (NLM) quality control (QC) review criteria. It is the responsibility of the sponsor or investigator to ensure that the study record is consistent with the NLM QC review criteria.
December 16, 2019

Last Verified

Last Verified

The most recent date on which the study sponsor or investigator confirmed the information about a clinical study on ClinicalTrials.gov as accurate and current. If a study with a recruitment status of recruiting; not yet recruiting; or active, not recruiting has not been confirmed within the past 2 years, the study's recruitment status is shown as unknown.
October 1, 2019

More Information

Terms related to this study

Keywords

Additional Relevant MeSH Terms

Other Study ID Numbers

Other Study ID Numbers

Identifiers or ID numbers other than the NCT number that are assigned to a clinical study by the study's sponsor, funders, or others. These numbers may include unique identifiers from other trial registries and National Institutes of Health grant numbers.
  • Yunwei Wei 2019-10-20

Plan for Individual participant data (IPD)

Plan to Share Individual Participant Data (IPD)?

NO

IPD Plan Description

Personal privacy involved

Drug and device information, study documents

Studies a U.S. FDA-regulated drug product

No

Studies a U.S. FDA-regulated device product

No

product manufactured in and exported from the U.S.

No

This information was retrieved directly from the website clinicaltrials.gov without any changes. If you have any requests to change, remove or update your study details, please contact register@clinicaltrials.gov. As soon as a change is implemented on clinicaltrials.gov, this will be updated automatically on our website as well.

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