Dataset Information

Measuring kinetics and metastatic propensity of CTCs by blood exchange between mice.

ABSTRACT: Existing preclinical methods for acquiring dissemination kinetics of rare circulating tumor cells (CTCs) en route to forming metastases have not been capable of providing a direct measure of CTC intravasation rate and subsequent half-life in the circulation. Here, we demonstrate an approach for measuring endogenous CTC kinetics by continuously exchanging CTC-containing blood over several hours between un-anesthetized, tumor-bearing mice and healthy, tumor-free counterparts. By tracking CTC transfer rates, we extrapolated half-life times in the circulation of between 40 and 260 s and intravasation rates between 60 and 107,000 CTCs/hour in mouse models of small-cell lung cancer (SCLC), pancreatic ductal adenocarcinoma (PDAC), and non-small cell lung cancer (NSCLC). Additionally, direct transfer of only 1-2% of daily-shed CTCs using our blood-exchange technique from late-stage, SCLC-bearing mice generated macrometastases in healthy recipient mice. We envision that our technique will help further elucidate the role of CTCs and the rate-limiting steps in metastasis.

SUBMITTER: Hamza B

PROVIDER: S-EPMC8479082 | biostudies-literature | 2021 Sep

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Measuring kinetics and metastatic propensity of CTCs by blood exchange between mice.

Hamza Bashar B Miller Alex B AB Meier Lara L Stockslager Max M Ng Sheng Rong SR King Emily M EM Lin Lin L DeGouveia Kelsey L KL Mulugeta Nolawit N Calistri Nicholas L NL Strouf Haley H Bray Christina C Rodriguez Felicia F Freed-Pastor William A WA Chin Christopher R CR Jaramillo Grissel C GC Burger Megan L ML Weinberg Robert A RA Shalek Alex K AK Jacks Tyler T Manalis Scott R SR

Nature communications 20210928 1

Existing preclinical methods for acquiring dissemination kinetics of rare circulating tumor cells (CTCs) en route to forming metastases have not been capable of providing a direct measure of CTC intravasation rate and subsequent half-life in the circulation. Here, we demonstrate an approach for measuring endogenous CTC kinetics by continuously exchanging CTC-containing blood over several hours between un-anesthetized, tumor-bearing mice and healthy, tumor-free counterparts. By tracking CTC trans ...[more]

PMID: 34584084

Similar Datasets

Project description:BackgroundEnumeration of circulating tumor cells (CTCs) has proven clinical significance for monitoring patients with metastatic cancers. Multiplexed gene expression profiling of CTCs is a potential tool for assessing disease status and monitoring treatment response. The Parsortix® technology enables the capture and harvest of CTCs from blood based on cell size and deformability. The HyCEAD™ (Hybrid Capture Enrichment Amplification and Detection) assay enables simultaneous amplification of short amplicons for up to 100 mRNA targets, and the Ziplex™ instrument quantifies the amplicons for highly sensitive gene expression profiling down to single cell levels. The aim of the study was to functionally assess this system.MethodsThe HyCEAD/Ziplex platform was used to quantify the expression levels for 72 genes using as little as 20 pg of total RNA or a single cultured tumor cell. Assay performance was evaluated using cells or total RNA spiked into Parsortix harvests of healthy donor blood. The assay was also evaluated using total RNA obtained from Parsortix harvests of blood from metastatic breast cancer (MBC) patients or healthy volunteers (HVs).ResultsUsing genes with low expression in WBC RNA and/or in unspiked Parsortix harvests from HVs, the assay distinguished between the different breast cancer and ovarian cancer cell lines with as little as 20 pg of total RNA (equivalent to a single cell) in the presence of 1 ng of WBC RNA. Single cultured cells spiked into Parsortix harvests from 10 mL of HV blood were also detected and distinguished from each other. CVs from repeatability experiments were less than 20%. Hierarchical clustering of clinical samples differentiated most MBC patients from HVs.ConclusionHyCEAD/Ziplex provided sensitive quantification of expression of 72 genes from 20 pg of total RNA from cultured tumor cell lines or from single cultured tumor cells spiked into lysates from Parsortix harvests of HV blood. The HyCEAD/Ziplex platform enables the quantification of selected genes in the presence of residual nucleated blood cells in Parsortix harvests. The HyCEAD/Ziplex platform is an effective tool for multiplexed molecular characterization of mRNA in small numbers of tumor cells harvested from blood.

Project description:BackgroundThe decline of photosynthesis in plants under low sink demand is well known. Previous studies focused on the relationship between stomatal conductance (gs) and net photosynthetic rate (Pn). These studies investigated the effect of changes in Photosystem II (PSII) function on the Pn decline under low sink demand. However, little is known about its effects on different limiting steps of electron transport chain in PSII under this condition.Methodology/principal findingTwo-month-old bean plants were processed by removing pods and flowers (low sink demand). On the 1(st) day after low sink demand treatment, a decline of Pn was accompanied by a decrease in gs and internal-to-ambient CO2 concentration ratio (Ci/Ca). From the 3(rd) to 9(th) day, Pn and gs declined continuously while Ci/Ca ratio remained stable in the treatment. Moreover, these values were lower than that of control. Wk (a parameter reflecting the damage to oxygen evolving complex of the donor side of PSII) values in the treatment were significantly higher than their corresponding control values. However, RCQA (a parameter reflecting the number of active RCs per excited cross-section of PSII) values in the treatment were significantly lower than control from the 5(th) day. From the 11(th) to 21(st) day, Pn and gs of the treatment continued to decline and were lower than control. This was accompanied by a decrease of RCQA, and an increase of Wk. Furthermore, the quantum yield parameters ?Po, ?Eo and ?Eo in the treatment were lower than in control; however, Ci/Ca values in the treatment gradually increased and were significantly higher than control on the 21(st) day.ConclusionsStomatal limitation during the early stage, whereas a combination of stomatal and non-stomatal limitation during the middle stage might be responsible for the reduction of Pn under low sink demand. Non-stomatal limitation during the late stages after the removal of the sink of roots and pods may also cause Pn reduction. The non-stomatal limitation was associated with the inhibition of PSII electron transport chain. Our data suggests that the donor side of PSII was the most sensitive to low sink demand followed by the reaction center of PSII. The acceptor side of PSII may be the least sensitive.

Dataset Information

Measuring kinetics and metastatic propensity of CTCs by blood exchange between mice.

Publications

Measuring kinetics and metastatic propensity of CTCs by blood exchange between mice.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets