Current strategies employed in the manipulation of gene expression for clinical purposes

Tsai, Hsing-Chuan; Pietrobon, Violena; Peng, Maoyu; Wang, Suning; Zhao, Lihong; Marincola, Francesco M.; Cai, Qi

doi:10.1186/s12967-022-03747-3

Cited by 17 publications

(10 citation statements)

References 300 publications

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“…Abnormal gene expression level or expression of genes containing deleterious variants can be at the basis of these genetic diseases. Currently, several molecular tools have been developed and are in clinical trials for therapeutic purposes acting to restore the physiological gene expression [ 70 ]. Thus, the whole knowledge of the transcription regulation mechanisms involving PRDM1 and PRDM2 genes in the immune biology can provide the molecular bases for the application of these new strategies to the autoimmune diseases.…”

Section: Discussionmentioning

confidence: 99%

Exploring the putative role of PRDM1 and PRDM2 transcripts as mediators of T lymphocyte activation

et al. 2023

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Background T cell activation and programming from their naïve/resting state, characterized by widespread modifications in chromatin accessibility triggering extensive changes in transcriptional programs, is orchestrated by several cytokines and transcription regulators. PRDM1 and PRDM2 encode for proteins with PR/SET and zinc finger domains that control several biological processes, including cell differentiation, through epigenetic regulation of gene expression. Different transcripts leading to main protein isoforms with (PR +) or without (PR-) the PR/SET domain have been described. Although many studies have established the critical PRDM1 role in hematopoietic cell differentiation, maintenance and/or function, the single transcript contribution has not been investigated before. Otherwise, very few evidence is currently available on PRDM2. Here, we aimed to analyze the role of PRDM1 and PRDM2 different transcripts as mediators of T lymphocyte activation. Methods We analyzed the transcription signature of the main variants from PRDM1 (BLIMP1a and BLIMP1b) and PRDM2 (RIZ1 and RIZ2) genes, in human T lymphocytes and Jurkat cells overexpressing PRDM2 cDNAs following activation through different signals. Results T lymphocyte activation induced an early increase of RIZ2 and RIZ1 followed by BLIMP1b increase and finally by BLIMP1a increase. The “first” and the “second” signals shifted the balance towards the PR- forms for both genes. Interestingly, the PI3K signaling pathway modulated the RIZ1/RIZ2 ratio in favor of RIZ1 while the balance versus RIZ2 was promoted by MAPK pathway. Cytokines mediating different Jak/Stat signaling pathways (third signal) early modulated the expression of PRDM1 and PRDM2 and the relationship of their different transcripts confirming the early increase of the PR- transcripts. Different responses of T cell subpopulations were also observed. Jurkat cells showed that the acute transient RIZ2 increase promoted the balancing of PRDM1 forms towards BLIMP1b. The stable forced expression of RIZ1 or RIZ2 induced a significant variation in the expression of key transcription factors involved in T lymphocyte differentiation. The BLIMP1a/b balance shifted in favor of BLIMP1a in RIZ1-overexpressing cells and of BLIMP1b in RIZ2-overexpressing cells. Conclusions This study provides the first characterization of PRDM2 in T-lymphocyte activation/differentiation and novel insights on PRDM1 and PRDM2 transcription regulation during initial activation phases.

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Section: Discussionmentioning

confidence: 99%

Exploring the putative role of PRDM1 and PRDM2 transcripts as mediators of T lymphocyte activation

et al. 2023

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“…This platform targets the regulatory region of endogenous genes through epigenetic modulation without any transient or permanent architectural changes of genomic DNA. This contrasts with the traditional genome-editing tools, including CRISPR, TALENS and ZFNs, which involves double strand break and irreversible chromosomal editing [ 9 ]. Moreover, the CRISPR activation platform allows simultaneous targeting of several endogenous genes with multiple sgRNAs cloned in the same vector.…”

Section: Discussionmentioning

confidence: 99%

“…However, genome-editing nucleases such as TALENs, zinc finger nucleases (ZFN) and clustered regularly interspaced short palindromic repeats (CRISPR)-associated systems (Cas) present their own limitations including permanent genomic alterations such as gross chromosomal rearrangements, a natural byproduct of simultaneous double strand breaks. The problem is compounded by multiple gene editing required when several functions need to be modulated simultaneously [ 9 ]. Therefore, a non-gene editing inducible system leading to transient and context-dependent modulation of cellular functions is preferable.…”

Section: Introductionmentioning

confidence: 99%

Nanoscale, antigen encounter-dependent, IL-12 delivery by CAR T cells plus PD-L1 blockade for cancer treatment

et al. 2023

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Background Chimeric antigen receptor (CAR)-T cell therapies for the treatment of hematological malignancies experienced tremendous progress in the last decade. However, essential limitations need to be addressed to further improve efficacy and reduce toxicity to assure CAR-T cell persistence, trafficking to the tumor site, resistance to an hostile tumor microenvironment (TME), and containment of toxicity restricting production of powerful but potentially toxic bioproducts to the TME; the last could be achieved through contextual release upon tumor antigen encounter of factors capable of converting an immune suppressive TME into one conducive to immune rejection. Methods We created an HER2-targeting CAR-T (RB-312) using a clustered regularly interspaced short palindromic repeats (CRISPR) activation (CRISPRa) system, which induces the expression of the IL-12 heterodimer via conditional transcription of its two endogenous subunits p35 and p40. This circuit includes two lentiviral constructs. The first one (HER2-TEV) expresses an anti-human epidermal growth factor receptor 2 (HER2) CAR single chain variable fragment (scFv), with CD28 and CD3z co-stimulatory domains linked to the tobacco etch virus (TEV) protease and two single guide RNAs (sgRNA) targeting the interleukin (IL)-12A and IL12B transcription start site (TSS), respectively. The second construct (LdCV) encodes linker for activation of T cells (LAT) fused to nuclease-deactivated Streptococcus Pyogenes Cas9 (dCas9)-VP64-p65-Rta (VPR) via a TEV-cleavable sequence (TCS). Activation of the CAR brings HER2-TEV in close proximity to LdCV releasing dCas9 for nuclear localization. This conditional circuit leads to conditional and reversible induction of the IL-12/p70 heterodimer. RB-312 was compared in vitro to controls (cRB-312), lacking the IL-12 sgRNAs and conventional HER2 CAR (convCAR). Results The inducible CRISPRa system activated endogenous IL-12 expression resulting in enhanced secondary interferon (FN)-γ production, cytotoxicity, and CAR-T proliferation in vitro, prolonged in vivo persistence and greater suppression of HER2+ FaDu oropharyngeal cancer cell growth compared to the conventional CAR-T cell product. No systemic IL-12 was detected in the peripheral circulation. Moreover, the combination with programmed death ligand (PD-L1) blockade demonstrated robust synergistic effects. Conclusions RB-312, the first clinically relevant product incorporating a CRISPRa system with non-gene editing and reversible upregulation of endogenous gene expression that promotes CAR-T cells persistence and effectiveness against HER2-expressing tumors. The autocrine effects of reversible, nanoscale IL-12 production limits the risk of off-tumor leakage and systemic toxicity.

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“…These systems are composed of sleeping beauty (SB) subsets and the PiggyBac (PB) [ 201 ]. For instance, the generation of transposon-engineered CAR-NK cells, was found to be effective to achieve stable expression without viral integration in addition to other advantages including capacity for large gene fragment transduction, increased biosafety, low immunogenicity and worthful cost-effectiveness [ 215 ].…”

Section: Car-nk Cell Therapy In Solid Tumors: Applications Challenges...mentioning

confidence: 99%

CAR-cell therapy in the era of solid tumor treatment: current challenges and emerging therapeutic advances

et al. 2023

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In the last decade, Chimeric Antigen Receptor (CAR)-T cell therapy has emerged as a promising immunotherapeutic approach to fight cancers. This approach consists of genetically engineered immune cells expressing a surface receptor, called CAR, that specifically targets antigens expressed on the surface of tumor cells. In hematological malignancies like leukemias, myeloma, and non-Hodgkin B-cell lymphomas, adoptive CAR-T cell therapy has shown efficacy in treating chemotherapy refractory patients. However, the value of this therapy remains inconclusive in the context of solid tumors and is restrained by several obstacles including limited tumor trafficking and infiltration, the presence of an immunosuppressive tumor microenvironment, as well as adverse events associated with such therapy. Recently, CAR-Natural Killer (CAR-NK) and CAR-macrophages (CAR-M) were introduced as a complement/alternative to CAR-T cell therapy for solid tumors. CAR-NK cells could be a favorable substitute for CAR-T cells since they do not require HLA compatibility and have limited toxicity. Additionally, CAR-NK cells might be generated in large scale from several sources which would suggest them as promising off-the-shelf product. CAR-M immunotherapy with its capabilities of phagocytosis, tumor-antigen presentation, and broad tumor infiltration, is currently being investigated. Here, we discuss the emerging role of CAR-T, CAR-NK, and CAR-M cells in solid tumors. We also highlight the advantages and drawbacks of CAR-NK and CAR-M cells compared to CAR-T cells. Finally, we suggest prospective solutions such as potential combination therapies to enhance the efficacy of CAR-cells immunotherapy.

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Current strategies employed in the manipulation of gene expression for clinical purposes

Cited by 17 publications

References 300 publications

Exploring the putative role of PRDM1 and PRDM2 transcripts as mediators of T lymphocyte activation

Exploring the putative role of PRDM1 and PRDM2 transcripts as mediators of T lymphocyte activation

Nanoscale, antigen encounter-dependent, IL-12 delivery by CAR T cells plus PD-L1 blockade for cancer treatment

CAR-cell therapy in the era of solid tumor treatment: current challenges and emerging therapeutic advances

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