Increasing the safety and efficacy of chimeric antigen receptor T cell therapy

Chimeric antigen receptor (CAR) T cell therapy is a promising cancer treatment that has recently been undergoing rapid development. However, there are still some major challenges, including precise tumor targeting to avoid off-target or "on-target/off-tumor" toxicity, adequate T cell infiltration and migration to solid tumors and T cell proliferation and persistence across the physical and biochemical barriers of solid tumors. In this review, we focus on the primary challenges and strategies to design safe and effective CAR T cells, including using novel cutting-edge technologies for CAR and vector designs to increase both the safety and efficacy, further T cell modification to overcome the tumor-associated immune suppression, and using gene editing technologies to generate universal CAR T cells. All these efforts promote the development and evolution of CAR T cell therapy and move toward our ultimate goal-curing cancer with high safety, high efficacy, and low cost.

Advancing chimeric antigen receptor T cell therapy with CRISPR/Cas9

The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated 9 (CRISPR/Cas9) system, an RNA-guided DNA targeting technology, is triggering a revolution in the field of biology. CRISPR/Cas9 has demonstrated great potential for genetic manipulation. In this review, we discuss the current development of CRISPR/Cas9 technologies for therapeutic applications, especially chimeric antigen receptor (CAR) T cell-based adoptive immunotherapy. Different methods used to facilitate efficient CRISPR delivery and gene editing in T cells are compared. The potential of genetic manipulation using CRISPR/Cas9 system to generate universal CAR T cells and potent T cells that are resistant to exhaustion and inhibition is explored. We also address the safety concerns associated with the use of CRISPR/Cas9 gene editing and provide potential solutions and future directions of CRISPR application in the field of CAR T cell immunotherapy. As an integration-free gene insertion method, CRISPR/Cas9 holds great promise as an efficient gene knock-in platform. Given the tremendous progress that has been made in the past few years, we believe that the CRISPR/Cas9 technology holds immense promise for advancing immunotherapy.

Novel T cells with improved in vivo anti-tumor activity generated by RNA electroporation

The generation of T cells with maximal anti-tumor activities will significantly impact the field of T-cell-based adoptive immunotherapy. In this report, we found that OKT3/IL-2-stimulated T cells were phenotypically more heterogeneous, with enhanced anti-tumor activity in vitro and when locally administered in a solid tumor mouse model. To further improve the OKT3/IL-2-based T cell manufacturing procedure, we developed a novel T cell stimulation and expansion method in which peripheral blood mononuclear cells were electroporated with mRNA encoding a chimeric membrane protein consisting of a single-chain variable fragment against CD3 and the intracellular domains of CD28 and 4-1BB (OKT3-28BB). The expanded T cells were phenotypically and functionally similar to T cells expanded by OKT3/IL-2. Moreover, co-electroporation of CD86 and 4-1BBL could further change the phenotype and enhance the in vivo anti-tumor activity. Although T cells expanded by the co-electroporation of OKT3-28BB with CD86 and 4-1BBL showed an increased central memory phenotype, the T cells still maintained tumor lytic activities as potent as those of OKT3/IL-2 or OKT3-28BB-stimulated T cells. In different tumor mouse models, T cells expanded by OKT3-28BB RNA electroporation showed anti-tumor activities superior to those of OKT3/IL-2 T cells. Hence, T cells with both a less differentiated phenotype and potent tumor killing ability can be generated by RNA electroporation, and this T cell manufacturing procedure can be further optimized by simply co-delivering other splices of RNA, thus providing a simple and cost-effective method for generating high-quality T cells for adoptive immunotherapy.

Application of gene therapy in tumor adoptive immunotherapy / 生物医学工程学杂志

Adoptive cell transfer of tumor-infiltrating lymphocyte (TIL) has resulted in clear and reproducible responses in a substantial percentage (approximately 50%) of patients with metastatic melanoma. The availability of tumor reactive TIL limits the use of adoptive cell transfer for the treatment of most non-melanoma cancer patients. Recent report indicated that adoptive transfer of T lymphocytes genetically modified with T-cell receptor (TCR) against a tumor antigen resulted in objective response in melanoma patients, thus shedding light on the use of this strategy for the treatment of common epithelial cancers beyond melanoma. In this review, the current status and potential use of genetic modification in the adoptive immunotherapy of cancer patients are be discussed.

Biopsy of needle stereotaction under mammography in nonpalpable breast lesion / 临床外科杂志

Objective To discuss the feasibility of diagnosis with needle stereotaction marking by steel wire under mammography and biopsy for nonpalpable early stage breast carcinoma.Methods Eighteen patients with doubtful nidus of breast only in X gram were performed needle stereotaction marking by steel wire under mammography, and removal freemartin biopsies of breast nidus were performed then.Results Localization and biopsy were very successful in all 18 patients. Seven patients were definited to have early stage breast carcinomas and eleven benign diseases.The diagnostic rate for nonpalpable early stage breast carcinoma was 100%.Conclusions Needle stereotaction marking by steel wire undermammography finish off the problem locating accurately breast nidus during operation.It was a credible and perfect technique for diagnoses of nonpalpable early stage breast carcinoma.

Surgical treatment of breast carcinoma / 中国普通外科杂志

Objective To explore the strategy of surgical therapy for breast carcinoma . Methods The clinical data of 258 patients with breast carcinoma were analysed retrospectively. Results (1)136 patients with stage Ⅰ～Ⅱ breast carcinoma were subjected to modified radical mastectomy, overall survival(OS) was 100%, and relapse free survival(RFS)92.6%.(2)Partial mastectomy and axillary dissection were performed on two patients with stage Ⅱbreast carcinoma,one relapsed in 5 months after operation. (3)In patients with stage Ⅲ breast carcinoma,there was no statistical difference in OS and RFS between 88 patients subjected to modified radical mastectomy and 20 radical mastectomy.(4)The radical operation showed a better efficacy in 5 patients with stage Ⅳ breast carcinoma.(5)Using special breast cutter and electrotome,the rate of surgical blood transfusion was 3.5%,postoperative hematocele 2.7%,flap necrosis 7.4%, effusion under skin 18.6%,and edema of affected limb 4.3%. Conclusions (1)The modified radical mastectomy is the major operation for stage Ⅰ～Ⅲ breast carcinoma patients. (2)Using special breast cutter and electrotome could cut down surgical blood transfusion and operation time.(3)Rational axillary lymph node dissective could reduce postoperative complications.

Diagnostic value of needle stereotaction marking by steel wire under mammography for nonpalpable breast carcinoma / 中国普通外科杂志

Objective To explore the diagnostic value of needle stereotaction marking by steel wire(NSMSW) under mammography for nonpalpable early stage breast carcinoma (BC). Methods 29 patients with nonpalpable breast lesions were performed NSMSW under mammography,and the lesions were removed for biopsy to make the diagnosis. Results Nine patients(31.0%) were definited as early stage BC and twenty(69.0%) benign disease.The pathologic results in freezing sections and in parafin sections were the same. Conclusions NSMSW under mammography can resolve the problem of accurate location of nonpalpable breast lesions during operation .It is a credible, useful and practical method for diagnosis of nonpalpable early stage BC.

Surgical therapy for breast intraductal papilloma / 中国普通外科杂志

Objective To study the effective therapy of breast intraductal papilloma . Method One hundred and thirty-two patients with breast intraductal papilloma (four patients with cancerization)were treated by resection of the tissue stained by methylene blue. The effect was evaluated. Results The pathological diagnosis were 91 patients with breast intraductal papilloma and 41 breast intraductal papillomatosis, and 4 of them were cancerization. One hundred and ninteen patients(90.1%) were followed up for 3～46 months, all patients were cured without recurrence after operation. Conclusions Resection of the tissue stained by methylene blue in treating breast intraductal papilloma is a reliable and effective method.