Table_1_Revolutionizing cancer immunotherapy in solid tumor: CAR engineering and single-cell sequencing insights.docx

The global increase in cancer incidence presents significant economic and societal challenges. While chimeric antigen receptor-modified T cell (CAR-T) therapy has demonstrated remarkable success in hematologic malignancies and has earned FDA approval, its translation to solid tumors encounters faces significant obstacles, primarily centered around identifying reliable tumor-associated antigens and navigating the complexities of the tumor microenvironment. Recent developments in single-cell RNA sequencing (scRNA-seq) have greatly enhanced our understanding of tumors by offering high-resolution, unbiased analysis of cellular heterogeneity and molecular patterns. These technologies have revolutionized our comprehension of tumor immunology and have led to notable progress in cancer immunotherapy. This mini-review explores the progress of chimeric antigen receptor (CAR) cell therapy in solid tumor treatment and the application of scRNA-seq at various stages following the administration of CAR cell products into the body. The advantages of scRNA-seq are poised to further advance the investigation of the biological characteristics of CAR cells in vivo, tumor immune evasion, the impact of different cellular components on clinical efficacy, the development of clinically relevant biomarkers, and the creation of new targeted drugs and combination therapy approaches. The integration of scRNA-seq with CAR therapy represents a promising avenue for future innovations in cancer immunotherapy. This synergy holds the potential to enhance the precision and efficacy of CAR cell therapies while expanding their applications to a broader range of malignancies.</p

Table_1_Single-cell RNA analysis to identify five cytokines signaling in immune-related genes for melanoma survival prognosis.xlsx

Melanoma is one of the deadliest skin cancers. Recently, developed single-cell sequencing has revealed fresh insights into melanoma. Cytokine signaling in the immune system is crucial for tumor development in melanoma. To evaluate melanoma patient diagnosis and treatment, the prediction value of cytokine signaling in immune-related genes (CSIRGs) is needed. In this study, the machine learning method of least absolute selection and shrinkage operator (LASSO) regression was used to establish a CSIRG prognostic signature of melanoma at the single-cell level. We discovered a 5-CSIRG signature that was substantially related to the overall survival of melanoma patients. We also constructed a nomogram that combined CSIRGs and clinical features. Overall survival of melanoma patients can be consistently predicted with good performance as well as accuracy by both the 5-CSIRG signature and nomograms. We compared the melanoma patients in the CSIRG high- and low-risk groups in terms of tumor mutation burden, infiltration of the immune system, and gene enrichment. High CSIRG-risk patients had a lower tumor mutational burden than low CSIRG-risk patients. The CSIRG high-risk patients had a higher infiltration of monocytes. Signaling pathways including oxidative phosphorylation, DNA replication, and aminoacyl tRNA biosynthesis were enriched in the high-risk group. For the first time, we constructed and validated a machine-learning model by single-cell RNA-sequencing datasets that have the potential to be a novel treatment target and might serve as a prognostic biomarker panel for melanoma. The 5-CSIRG signature may assist in predicting melanoma patient prognosis, biological characteristics, and appropriate therapy.</p

Residual drug volumes after infection of JAR cells with V-BCRPi according to flow cytometry analysis.

<p>The results of experimental groups 1 to 9 are shown: 1: mock cells, 2: cells with Mit, 3: cells with Mit and Ko143, or cells with Mit and infected with 4: V-BCRP1i, 5: V-BCRP2i, 6: V-BCRP3i, 7: V-BCRP1i-c, 8: V-BCRP2i-c, or 9: V-BCRP3i-c. Each residual drug volume represents the mean value of three independent experiments. *<i>P</i><0.01.</p

<i>DAX-1</i> mutations and SNPs identified in the secretory azoospermia patients and controls.

<p><i>DAX-1</i> mutations and SNPs identified in the secretory azoospermia patients and controls.</p

Changes in the apoptosis rate of transplantation tumors in nude mice injected with virus and 5-FU (x±s, n = 5).

<p>*: PBS+5-FU vs. V-BCRPi+5-FU, p<0.01.</p

MOESM3 of In vitro chemokine (C-C motif) receptor 6-dependent non-inflammatory chemotaxis during spermatogenesis

Additional file 3: Figure S3. The involvement of CCR6 in the chemotaxis of mouse spermatogenic cell lines (GC-1 and -2) induced by rDEFB1 or primary sertoli cells in vitro. The chemotactic activity of rDEFB1 to GC-1 (A) and GC-2 (B), with or without the immunodepletion of anti-CCR6 antibody. The chemotactic activity of the culture supernatant of primary Sertoli cells to GC-1 (C) and GC-2 (D), with or without the immunodepletion of anti-CCR6 antibody. *p < 0.05, **p < 0.01 and ***p < 0.001. Normal rabbit IgG was used as negative control. n = 10 in each group

List of missense mutations predicted by PolyPhen 2.0 and Mutation Taster.

<p>List of missense mutations predicted by PolyPhen 2.0 and Mutation Taster.</p

Knockdown of BCRP/ABCG2 expression by V-BCRPi in tumor cell bodies, as shown by Western blot analysis.

<p>1: Tumor body injected with PBS alone; 2: Tumor body injected with PBS and 5-FU; 3: Tumor body injected with V-BCRPi alone; and 4: Tumor body injected with V-BCRPi and 5-FU. It was concluded from the experimental results that the expression of <i>BCRP/ABCG2</i> in tumors injected with V-BCRPi (with 5-FU treatment) was lower than that of the un-injected tumors (with PBS and 5-FU). There was no difference among the groups injected with the various V-BCRPi retroviruses and 5-FU.</p

The effect of the DAX-1 V385L variant on AR function.

<p>A WT or mutant DAX-1 expression vector was cotransfected with an AR expression vector and a testosterone-inducible pMMTV-LUC plasmid into HeLa cells, and luciferase activity was measured with or without testosterone treatment. Compared with the WT and the other mutants, the DAX-1 V385L mutant failed to repress the transcriptional activity of AR in the presence of testosterone. The results are presented as the fold-change of testosterone treated relative to that in the vehicle-treated control. NC: pcDNA3.1-HA were cotransfected with an AR expression vector and a testosterone-inducible pMMTV-LUC plasmid into HeLa cells. (* <i>p</i> < 0.05).</p

Changes in tumor weight and volume and the anti-tumor rate in transplantation tumors in nude mice after virus infection and drug treatment (x±s, n = 15).

<p>*: PBS+5-FU vs. V-BCRPi+5-FU, p<0.01.</p