Transcriptome Reprogramming of CD11b+ Bone Marrow Cells by Pancreatic Cancer Extracellular Vesicles

Maia, Joana; Otake, Andréia Hanada; Poças, Juliana; Carvalho, Ana Sofía; Beck, Hans Christian; Magalhães, Ana; Matthiesen, Rune; Moraes, Maria Carolina Strano; Costa-Silva, Bruno

doi:10.3389/fcell.2020.592518

Cited by 13 publications

(11 citation statements)

References 106 publications

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“…Most therapeutic antibodies that are currently available are developed in the IgG framework; the majority of which belong to the IgG1 subtype [ 67 , 68 ]. IgG1 has a 60% abundance distribution in the serum and primarily binds protein antigens, is highly stable and resistant to aggregation, and has a greater ability to activate antibody-dependent cytotoxicity and complement-dependent cytotoxicity [ 69 , 70 ]. In contrast, another antibody fragment based on the IgG form, scFv-Fc, is similar to full-length IgG in terms of tumor uptake [ 66 ] and has a better tumor targeting profile.…”

Section: Discussionmentioning

confidence: 99%

Fully human recombinant antibodies against EphA2 from a multi-tumor patient immune library suitable for tumor-targeted therapy

Yang

Nian

et al. 2021

Bioengineered

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Enhanced EphA2 expression is observed in a variety of epithelial-derived malignancies and is an important target for anti-tumor therapy. Currently, Therapeutic monoclonal antibodies against immune checkpoints have shown good efficacy for tumor treatment. In this study, we constructed an immune single-chain fragment variable (scFv) library using peripheral blood mononuclear cells (PBMCs) from 200 patients with a variety of malignant tumors. High affinity scFvs against EphA2 can be easily screened from the immune library using phage display technology. Anti-EphA2 scFvs can be modified into any form of recombinant antibody, including scFv-Fc and full-length IgG1 antibodies, and the recombinant antibody affinity was improved following modification. Among the modified anti-EphA2 antibodies the affinity of 77-IgG1 was significantly increased, reaching a pmol affinity level (10 −12 ). We further demonstrated the binding activity of recombinant antibodies to the EphA2 protein, tumor cells, and tumor tissues using macromolecular interaction techniques, flow cytometry and immunohistochemistry. Most importantly, both the constructed scFvs-Fc, as well as the IgG1 antibodies against EphA2 were able to inhibit the growth of tumor cells to some extent. These results suggest that the immune libraries from patients with malignant tumors are more likely to screen for antibodies with high affinity and therapeutic effect. The constructed fully human scFv immune library has broad application prospects for specific antibody screening. The screened scFv-Fc and IgG1 antibodies against EphA2 can be used for the further study of tumor immunotherapy.

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Section: Discussionmentioning

confidence: 99%

Fully human recombinant antibodies against EphA2 from a multi-tumor patient immune library suitable for tumor-targeted therapy

Yang

Nian

et al. 2021

Bioengineered

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“…Cells were maintained at 37 • C at 5% CO 2 levels. For conditioning, cells were cultured in the designated media supplemented with 1% penicillin-streptomycin and 10% EV-depleted FBS [26]. For the preparation of conditioned culture media (CCMs), cells were seeded in 150 mm culture dishes containing 20 mL of EVdepleted medium and grown for 72 h until they reached a confluency of ~90%.…”

Section: Methodsmentioning

confidence: 99%

Defining Optimal Conditions for Tumor Extracellular Vesicle DNA Extraction for Mutation Profiling

Elzanowska

Berrocal

García‐Peláez

et al. 2022

Cancers

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(1) Background: Extracellular vesicles (EVs) have emerged as crucial players in the communication between cells in both physiological and pathological scenarios. The functions of EVs are strongly determined by their molecular content, which includes all bioactive molecules, such as proteins, lipids, RNA, and, as more recently described, double-stranded DNA. It has been shown that in oncological settings DNA associated with EVs (EV-DNA) is representative of the genome of parental cells and that it reflects the mutational status of the tumor, gaining much attention as a promising source of biomarker mutant DNA. However, one of the challenges in studies of EV-DNA is the lack of standardization of protocols for the DNA extraction from EVs, as well as ways to assess quality control, which hinders its future implementation in clinics. (2) Methods: We performed a comprehensive comparison of commonly used approaches for EV-DNA extraction by assessing DNA quantity, quality, and suitability for downstream analyses. (3) Results: We here established strategic points to consider for EV-DNA preparation for mutational analyses, including qPCR and NGS. (4) Conclusions: We put in place a workflow that can be applied for the detection of clinically relevant mutations in the EV-DNA of cancer patients.

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“…Prior to analysis, plasma samples were aliquoted and stored at −80 °C. A protocol we previously described involving sequential ultracentrifugation combined with a sucrose cushion was used to purify the EVs [ 23 ].…”

Section: Methodsmentioning

confidence: 99%

IgG+ Extracellular Vesicles Measure Therapeutic Response in Advanced Pancreatic Cancer

Couto

Elzanowska

Maia

et al. 2022

Cells

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(1) Background: Pancreatic ductal adenocarcinoma (PDAC) is expected to be the second-leading cause of cancer deaths by 2030. Imaging techniques are the standard for monitoring the therapy response in PDAC, but these techniques have considerable limits, including delayed disease progression detection and difficulty in distinguishing benign from malignant lesions. Extracellular vesicle (EV) liquid biopsy is an emerging diagnosis modality. Nonetheless, the majority of research for EV-based diagnosis relies on point analyses of EVs at specified times, while longitudinal EV population studies before and during therapeutic interventions remain largely unexplored. (2) Methods: We analyzed plasma EV protein composition at diagnosis and throughout PDAC therapy. (3) Results: We found that IgG is linked with the diagnosis of PDAC and the patient’s response to therapy, and that the IgG+ EV population increases with disease progression and reduces with treatment response. Importantly, this covers PDAC patients devoid of the standard PDAC seric marker CA19.9 expression. We also observed that IgG is bound to EVs via the tumor antigen MAGE B1, and that this is independent of the patient’s inflammatory condition and IgG seric levels. (4) Conclusions: We here propose that a population analysis of IgG+ EVs in PDAC plasma represents a novel method to supplement the monitoring of the PDAC treatment response.

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Transcriptome Reprogramming of CD11b+ Bone Marrow Cells by Pancreatic Cancer Extracellular Vesicles

Cited by 13 publications

References 106 publications

Fully human recombinant antibodies against EphA2 from a multi-tumor patient immune library suitable for tumor-targeted therapy

Fully human recombinant antibodies against EphA2 from a multi-tumor patient immune library suitable for tumor-targeted therapy

Defining Optimal Conditions for Tumor Extracellular Vesicle DNA Extraction for Mutation Profiling

IgG+ Extracellular Vesicles Measure Therapeutic Response in Advanced Pancreatic Cancer

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