Method for improving anti-tumor effect of rituximab and application

Abstract

The invention belongs to the technical field of bioengineering, and discloses a method for improving the anti-tumor effect of rituximab and application. The method comprises the following steps that sgRNA and cas9mRNA of specific PIG-A are packaged through lipid nanoparticles; a CD20 antibody is coupled by utilizing a lipid nanoparticle transmission system, and B-NHL lymphoma cells and lymphoma cells PIG-A are accurately targeted; and by editing PIG-A of the B-NHL cells, synthesis of GPI in the cells is impaired, and proteins including extracellular ankyrin CD59 are lost. According to the method, through gene editing of PIG-A, synthesis of GPI and expression and distribution of ankyrin such as CD59 and CD55 are reduced, and drug sensitivity and tumor killing effect of rituximab can be greatly improved; and a lipid nanoparticle transmission system is used for coupling a CD20 antibody, so that rituximab drug-resistant B-NHL cells can kill 99% of lymphoma cells in initial treatment.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and in particular relates to a method and application for improving the anti-tumor effect of rituximab. Background technique [0002] At present, B-cell Non-Hodgkin's lymphoma (-NHL) accounts for about 70% of all NHLs, and most of them are aggressive and heterogeneous. The first manifestation is painless lymphadenopathy in the neck or supraclavicular, which can involve various organs in the whole body. The clinical manifestations are complex and the course of the disease varies in length. With the aging population in my country and the accumulation of risk factors such as chronic infection, unhealthy lifestyle, and environmental pollution, the incidence of B-NHL is increasing year by year, and the age of onset is constantly moving forward. The current standard CHOP (cyclophosphamide + doxorubicin + vinblastine + prednisone) regimen combined with human-mouse chimeric anti-CD20 monoclonal a...

AI Technical Summary

Problems solved by technology

[0008] (1) The 5-year survival rate of patients receiving first-line chemotherapy is 60%-70%, but more than 30% of patients will develop relapsed / refractory B-NHL due to resistance to targeted and chemotherapy drugs, resulting in Poor prognosis, many of whom show resistance to rituximab

[0009] (2) By increasing the dose of rituximab and replacing other regimens or even hematopoietic stem cell transplantation, it is still impossible to fundamentally solve the rituximab resistance and cure such patients

[0010] (3) None of the existing methods can fundamentally reverse rituximab drug resistance, and they do not have the targeting of B-lineage lymphoma cells

[0011] (4) The method of inducing PIG-A mutation in B-NHL cells to improve rituximab drug sensitivity has not been reported

[0012] (5) The existing CRISPR / Cas9 system is mostly packaged and transmitted by adenovirus, which is limited by the carrying capacity of adenovirus, and it is not possible to make too many modifications to PIG-A-CRISPR / Cas9 to improve the precise targeting of the system B lymphoma cells

[0014] 1. There is no standard treatment for relapsed and refractory B-NHL, and most of them enter clinical trials, switch to other chemotherapy drugs or high-dose chemotherapy / combined with incremental rituximab treatment, the curative effect is uncertain and the side effects after chemotherapy are relatively large. The risk of chemotherapy and the cost of treatment increase, and according to many studies, even if remission is achieved, the relapse rate is relatively high;

[0015] 2. The treatment of relapsed and refractory B-NHL uses new immunotherapy, CART cells and new monoclonal antibodies such as POLA monoclonal antibody. The existing CART cell therapy has indeed made encouraging progress, but the effect on lymphoma is still not satisfactory , mostly relapse within one year, the CD79 monoclonal antibody, namely POLA, is very expensive and has poor accessibility, and clinical data show that its maintenance time after remission is short;

[0016] 3. Gene editing technology is gradually applied in clinical practice, but for relapsed and refractory B-NHL, there is still no precise target, and the risk of off-target is relatively high;

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