Evaluation of Nonviral piggyBac and lentiviral Vector in Functions of CD19chimeric Antigen Receptor T Cells and Their Antitumor Activity for CD19+ Tumor Cells

Lin, Zhi; Liu, Xiangzhen; Liu, Tao; Gao, Huile; Wang, Sitong; Zhu, Xiaogang; Rong, Lijie; Cheng, Jingbo; Cai, Zhigang; Xu, Fuqiang; Tan, Xue; Lv, Linjie; Zhong, Li; Sun, Yan; Qian, Qijun

doi:10.3389/fimmu.2021.802705

Cited by 10 publications

(10 citation statements)

References 27 publications

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“…The discovery of transposons, capable of achieving genomic integration efficacy close to the viral systems (Lin et al, 2022) has given hope to patients who require lifelong gene expression without the risks associated with viral systems. The integration ability of the piggyBac transposon with the aid of piggyBac transposase to recognize ("cut") the inverted terminal repeats (ITR) and its integration ("paste") into the genome ensures long-term gene expression with nonrandom insertion.…”

Section: Discussionmentioning

confidence: 99%

An optimized polymeric delivery system for piggyBac transposition

Meenakshi Sundaram,

Bahadur K. C.,

et al. 2024

Biotech & Bioengineering

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The piggyBac transposon/transposase system has been explored for long‐term, stable gene expression to execute genomic integration of therapeutic genes, thus emerging as a strong alternative to viral transduction. Most studies with piggyBac transposition have employed physical methods for successful delivery of the necessary components of the piggyBac system into the cells. Very few studies have explored polymeric gene delivery systems. In this short communication, we report an effective delivery system based on low molecular polyethylenimine polymer with lipid substitution (PEI‐L) capable of delivering three components, (i) a piggyBac transposon plasmid DNA carrying a gene encoding green fluorescence protein (PB‐GFP), (ii) a piggyBac transposase plasmid DNA or mRNA, and (iii) a 2 kDa polyacrylic acid as additive for transfection enhancement, all in a single complex. We demonstrate an optimized formulation for stable GFP expression in two model cell lines, MDA‐MB‐231 and SUM149 recorded till day 108 (3.5 months) and day 43 (1.4 months), respectively, following a single treatment with very low cell number as starting material. Moreover, the stability of the transgene (GFP) expression mediated by piggyBac/PEI‐L transposition was retained following three consecutive cryopreservation cycles. The success of this study highlights the feasibility and potential of employing a polymeric delivery system to obtain piggyBac‐based stable expression of therapeutic genes.

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Section: Discussionmentioning

confidence: 99%

An optimized polymeric delivery system for piggyBac transposition

Meenakshi Sundaram,

Bahadur K. C.,

et al. 2024

Biotech & Bioengineering

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“…Lin et al. observed a higher level of distribution of anti-CD19 pbCAR T cells in mesenteric lymph nodes, bone marrow of the femur, spleen, kidneys, and lungs, specifically accumulating at CD19-rich sites and CD19-positive Raji cell-induced tumors ( 19 ). During the whole CAR T process, the patient did not feel any pain or discomfort in the abdominal cavity.…”

Section: Discussionmentioning

confidence: 99%

“…The CAR T cells could be detected on day 7 in the ascites, and the absolute value was higher than that in the blood, showing good penetration and infiltration of CAR T cells from peripheral blood to the peritoneum. Lin et al observed a higher level of distribution of anti-CD19 pbCAR T cells in mesenteric lymph nodes, bone marrow of the femur, spleen, kidneys, and lungs, specifically accumulating at CD19-rich sites and CD19positive Raji cell-induced tumors (19). During the whole CAR T process, the patient did not feel any pain or discomfort in the abdominal cavity.…”

Section: Figurementioning

confidence: 99%

Rapid response in relapsed follicular lymphoma with massive chylous ascites to anti-CD19 CAR T therapy using Piggy Bac: A case report

et al. 2022

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CD19-directed chimeric antigen receptor (CAR) T cell therapy has been shown to achieve a considerably durable response in patients with refractory or relapsed B cell non-Hodgkin lymphomas, as seen from the results of Zuma-1, Zuma-5, and other clinical trials. Most of these CARs were generated by lentivirus or reverse adenovirus. It is rare to see CARs using non-viral vectors, such as Piggy Bac (pb), in treating lymphoma patients with active diseases. Generally, patients with a high tumor burden tend to have a higher rate of severe cytokine release syndrome (CRS) or neurological events as reported in the literature. Patients with symptomatic pleural effusions are excluded from the Zuma-1 trial because of the risk of severe CRS. We report here that a patient with relapsed follicular lymphoma with bulky disease and massive chylous ascites failed several lines of chemotherapy. After infusion of the CD19-directed pbCAR-T cells at 6 × 106 cells/kg, the patient had a rapid response and achieved a nearly complete metabolic remission on day 28. There was only grade 1 CRS, and no neurotoxicity occurred. The CAR-T cells reached a peak level on day 14 and spread into the ascites and expanded for 3 months. This might be the first case reported for pbCAR-T cells to treat relapsed follicular lymphoma directly. The long-term efficacy will be observed, and more patients be tested in the future.Clinical Trial Registrationhttps://ClinicalTrials.gov, identifier NCT05472610.

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“…In addition, PB does not leave gene excision marks, so that possible genome damage is less likely. Transposition involving PB is also simple to reproduce in vitro [25]. Targeted CAR transgene insertion can also be performed using the CRISPR-Cas9 genome editing system.…”

Section: Production Of Car-t Cells and Application In Clinical Practicementioning

confidence: 99%

A living drug: application of CAR-T therapy for lymphoid malignancies and beyond

et al. 2022

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The ongoing development of novel personalized cancer therapies has resulted in the implementation of T-cells enriched with synthetic chimeric antigen receptors, known as chimeric antigen receptors T-cell (CAR-T) cells, into clinical practice. CAR-T cells are able to recognize and bind specific antigens present on the surface of target cellsso-called tumor-associated antigens. This innovative method has been approved for the treatment of hematological malignancies and may also serve as a bridge to hematopoietic stem cell transplantation. The production of the drug containing modified T-cells consists of several steps -leukapheresis, T-cell activation, transduction and expansion of the final CAR-T cells. Activation of CAR-T cells occurs through a pathway independent of the major histocompatibility complex, which is often associated with uncontrolled responses from the immune system and adverse reactions such as cytokine release syndrome. CAR-T therapy can only be performed in certified centers, and requires close cooperation between experienced specialists of different medical disciplines. This is what determines its effectiveness. Every step from collection and cryopreservation, through transport and modification, to thawing and infusion is strictly controlled because it has a critical impact on the quality and efficiency of the drug. Despite its proven benefits, CAR-T therapy remains available only to patients who meet well-defined criteria. These however are liable to change with the emergence of new indications.

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Evaluation of Nonviral piggyBac and lentiviral Vector in Functions of CD19chimeric Antigen Receptor T Cells and Their Antitumor Activity for CD19+ Tumor Cells

Cited by 10 publications

References 27 publications

An optimized polymeric delivery system for piggyBac transposition

An optimized polymeric delivery system for piggyBac transposition

Rapid response in relapsed follicular lymphoma with massive chylous ascites to anti-CD19 CAR T therapy using Piggy Bac: A case report

A living drug: application of CAR-T therapy for lymphoid malignancies and beyond

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