Posttransplantation Bone Marrow Assessment by Quantifying Hematopoietic Cell–Derived mRNAs in Plasma Exosomes/Microvesicles

Aoki, Jun; Ohashi, Kazuteru; Mitsuhashi, Masato; Murakami, Taku; Oakes, Melanie L.; Kobayashi, Takeshi; Kakihana, Kazuhiko; Sakamaki, Hisashi

doi:10.1373/clinchem.2013.213850

Cited by 23 publications

(21 citation statements)

References 16 publications

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“…As previously described. 34,35 In this system, cDNA primed by specific primers comes to solution, while oligo (dT)-primed cDNA remains immobilized on the plate. Reverse transcription PCR of the solubilized specifically primed cDNA follows.…”

Section: Flow Cytometrymentioning

confidence: 99%

“…In the absence of reverse transcriptase, no amplification takes place in this system. 34,35 For PCR, the initial temperature cycle of 95 C for 5 min was followed by 45 cycles of 95 C for 30 s, 65 C for 1 min in a final volume of 5 mL of 1x SYBR green PCR master mix (Bio-Rad, Hercules, CA, USA) in a 384-well plate. The cycle threshold (Ct) was determined by the analytical software (SDS, Applied Biosystems).…”

Section: Flow Cytometrymentioning

confidence: 99%

“…The sensitivity and reproducibility of this method was previously described. 34,35 Statistics The values for total exosomal protein determined for samples of off-protocol glioma patients, on-protocol patients at diagnosis and NC were compared using the two-tailed Wilcoxon test. Paired (pre-post therapy) clinical trial specimens were plotted by subject, and the pre and post values were compared using the two tailed signed rank test.…”

Section: Flow Cytometrymentioning

confidence: 99%

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Exosomes isolated from plasma of glioma patients enrolled in a vaccination trial reflect antitumor immune activity and might predict survival

et al. 2015

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Exosomes in plasma of glioma patients hold promise as biomarkers of prognosis. We aimed to determine whether changes in total exosomal protein and mRNA expression levels could serve as surrogate markers of immunological and clinical responses in glioma patients receiving antitumor vaccines. Exosomes were isolated from pre/post-vaccine plasma specimens in 20/22 patients enrolled in a phase I/II trial with the antitumor vaccine. Exosomal protein content was analyzed and mRNA expression levels for 24 genes were simultaneously assessed by qRT-PCR. Pre-to postvaccination changes in exosomal protein and DCt values were correlated with immunological and clinical responses and survival using Spearman rank statistics and hazard ratios (HR). Exosomal protein levels positively correlated (p < 0.0043) with the WHO tumor grade at diagnosis. Protein levels were lower in post-vs. pre-vaccination exosome fractions. Post-therapy increases in tumor size were associated with elevations in exosome proteins in glioblastoma but not always in anaplastic astrocytoma (AA). Only exosomal DCt values for IL-8, TIMP-1, TGF-b and ZAP70 were significant (p < 0.04 to p < 0.001). The DCt for IL-8 and TGF-b mRNA positively correlated with post-vaccine immunologic responses to glioma antigens, while DCt for TIMP-1 mRNA was negatively correlated to DCt for IL-8 and TGF-b. Only DCt for IL-8 weakly correlated with OS and time to progression (TTP). In post-vaccine exosomes of the longest surviving patient with AA, mRNA for PD-1 was persistently elevated. Protein and mRNA expression levels for immune-related genes in plasma exosomes were useful in evaluating glioma patients' response to vaccination therapy.

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Section: Flow Cytometrymentioning

confidence: 99%

Section: Flow Cytometrymentioning

confidence: 99%

Section: Flow Cytometrymentioning

confidence: 99%

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Exosomes isolated from plasma of glioma patients enrolled in a vaccination trial reflect antitumor immune activity and might predict survival

et al. 2015

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“…Furthermore, plasma EVs from HMs patients undergoing alloSCT contains myeloid, erythroid, and megakaryocyte lineage specific transcripts. Interestingly, the presence of these EV mRNAs seems to precede and predict the recovery of white blood cells, reticulocyte, and platelet count in blood, thus providing a novel potential biomarker of allogeneic stem cells engraftment [154]. …”

Section: Clinical Potential Of Evs As Biomarkers and Therapeuticsmentioning

confidence: 99%

Extracellular Vesicles in Hematological Malignancies: From Biology to Therapy

Caivano

Rocca

Laurenzana

et al. 2017

IJMS

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Extracellular vesicles (EVs) are a heterogeneous group of particles, between 15 nanometers and 10 microns in diameter, released by almost all cell types in physiological and pathological conditions, including tumors. EVs have recently emerged as particularly interesting informative vehicles, so that they could be considered a true “cell biopsy”. Indeed, EV cargo, including proteins, lipids, and nucleic acids, generally reflects the nature and status of the origin cells. In some cases, EVs are enriched of peculiar molecular cargo, thus suggesting at least a degree of specific cellular packaging. EVs are identified as important and critical players in intercellular communications in short and long distance interplays. Here, we examine the physiological role of EVs and their activity in cross-talk between bone marrow microenvironment and neoplastic cells in hematological malignancies (HMs). In these diseases, HM EVs can modify tumor and bone marrow microenvironment, making the latter “stronger” in supporting malignancy, inducing drug resistance, and suppressing the immune system. Moreover, EVs are abundant in biologic fluids and protect their molecular cargo against degradation. For these and other “natural” characteristics, EVs could be potential biomarkers in a context of HM liquid biopsy and therapeutic tools. These aspects will be also analyzed in this review.

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“…In the first scenario, HSCs could increase their proliferation rate within the standard active hematopoietic sites. 12 Alternatively, HSCs could recolonize intraosseous spaces that were abandoned because they were redundant for the standard homeostatic need, thus restoring the hematopoietic asset classically described in infancy. 13 These 2 different patterns would imply divergent signaling mechanisms, whose relative contribution is yet undefined, because of the obvious concerns in performing repeated BM biopsies in multiple bone segments.…”

Section: Introductionmentioning

confidence: 99%

Allogeneic cell transplant expands bone marrow distribution by colonizing previously abandoned areas: an FDG PET/CT analysis

Fiz

Marini

Campi

et al. 2015

Blood

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Posttransplantation Bone Marrow Assessment by Quantifying Hematopoietic Cell–Derived mRNAs in Plasma Exosomes/Microvesicles

Cited by 23 publications

References 16 publications

Exosomes isolated from plasma of glioma patients enrolled in a vaccination trial reflect antitumor immune activity and might predict survival

Exosomes isolated from plasma of glioma patients enrolled in a vaccination trial reflect antitumor immune activity and might predict survival

Extracellular Vesicles in Hematological Malignancies: From Biology to Therapy

Allogeneic cell transplant expands bone marrow distribution by colonizing previously abandoned areas: an FDG PET/CT analysis

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