PD-1 gene silenced CD133-targeting CAR T cell and application thereof

Abstract

The invention provides a PD-1 gene silenced CD133-targeting CAR T cell and application thereof. The PD-1 gene silenced CD133-targeting CAR T cell is characterized in that a preparation method thereofcomprises the following steps: firstly importing CRISPR-Cas9 and a transposon system into PBMC by using a nucleofection system, activating a T cell, and conducting T cell expansion to obtain the PD-1gene silenced CD133-targeting CAR T cell. The PD-1 gene silenced CD133-targeting CAR T cell provided by the invention can solve PD-1 pathway mediated immunosuppression and prevent tumor metastasis andrecurrence. In comparison with PD-1 or PD-L1 antibody combination therapy, the PD-1 gene silenced CD133-targeting CAR T cell can effectively avoid systemic toxicity problems and accurately regulate the immune microenvironment.

Description

technical field [0001] The invention belongs to the field of tumor immune cell therapy, and in particular relates to a CART cell targeting CD133 for PD-1 gene silencing and an application thereof. Background technique [0002] The cancer stem cell hypothesis states that there is a small group of cells in tumors that have the ability to self-proliferate, initiate tumorigenesis, and differentiate into other tumor cells. A large number of experimental data also revealed the existence of this group of cells. More importantly, studies have shown that cancer stem cells are resistant to drugs, which is related to the failure of traditional treatments such as chemotherapy and radiotherapy, and is an important factor leading to tumor metastasis and recurrence. Therefore, targeting cancer stem cells will be the key to the success of tumor therapy [1]. [0003] CD133 is currently one of the most commonly used molecular markers for identifying tumor stem cells. It is expressed in a va...

AI Technical Summary

Problems solved by technology

[0007] However, there are at least two problems in the use of immune checkpoint blocking antibodies for immunotherapy: 1. Immune checkpoint blocking antibodies will also break the normal immune tolerance of the body while treating tumors, causing normal tissues and organs of the body to be activated. Cell attack, long-term use will cause systemic toxicity [16]; 2. The immune checkpoint contains multiple members, and blocking the PD-1 / PD-L1 pathway may not be enough

However, it takes a long time and investment to develop multiple sets of highly efficient antibodies, so it is difficult to develop multiple antibodies in combination

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