A Global Federated Real-World Data and Analytics Platform for Research

Objective This article describes a scalable, performant, sustainable global network of electronic health record data for biomedical and clinical research. Materials and Methods TriNetX has created a technology platform characterized by a conservative security and governance model that facilitates collaboration and cooperation between industry participants, such as pharmaceutical companies and contract research organizations, and academic and community-based healthcare organizations (HCOs). HCOs participate on the network in return for access to a suite of analytics capabilities, large networks of de-identified data, and more sponsored trial opportunities. Industry participants provide the financial resources to support, expand, and improve the technology platform in return for access to network data, which provides increased efficiencies in clinical trial design and deployment. Results TriNetX is a growing global network, expanding from 55 HCOs and 7 countries in 2017 to over 220 HCOs and 30 countries in 2022. Over 19 000 sponsored clinical trial opportunities have been initiated through the TriNetX network. There have been over 350 peer-reviewed scientific publications based on the network’s data. Conclusions The continued growth of the TriNetX network and its yield of clinical trial collaborations and published studies indicates that this academic-industry structure is a safe, proven, sustainable path for building and maintaining research-centric data networks

Measurements of accuracy and precision of LiquidBiopsy capture of MCF7 and H1650 cells.

<p>Measurements of accuracy and precision of LiquidBiopsy capture of MCF7 and H1650 cells.</p

LiquidBiopsy automated platform.

<p>(A) Diagram of the LiquidBiopsy platform. (B) Closeup of the platform worksurface. A X-Y pipetting arm with 4 pipettor heads transfers sample, buffers and antibody stains into 4 flow cells in the manifold. Sheath buffers are controlled by pumps on the rear wall of the platform.</p

Validation of Automated CTC flow cell operation on LiquidBiopsy platform.

<p>(A) Plot of CTC recovery as a function of spike-in density for MCF7 and H1650 cells. (B) Plot of sample purity as a function of spike in density for MCF7 and H1650 cells. The orange dots indicate the predicted purity of MCF7 cells if the non target recovery is held constant at 55 cells/mL. (C) Raw data and linear fit to MCF7 recovery curve from 9 to 90 cells/mL. (D) Raw data and linear fit to H1650 recovery curve from 9 to 300 cells/mL.</p

Results of intra and inter assay precision studies.

<p>Results of intra and inter assay precision studies.</p

Circulating Tumor Cells: Clinically Relevant Molecular Access Based on a Novel CTC Flow Cell

<div><p>Background</p><p>Contemporary cancer diagnostics are becoming increasing reliant upon sophisticated new molecular methods for analyzing genetic information. Limiting the scope of these new technologies is the lack of adequate solid tumor tissue samples. Patients may present with tumors that are not accessible to biopsy or adequate for longitudinal monitoring. One attractive alternate source is cancer cells in the peripheral blood. These rare circulating tumor cells (CTC) require enrichment and isolation before molecular analysis can be performed. Current CTC platforms lack either the throughput or reliability to use in a clinical setting or they provide CTC samples at purities that restrict molecular access by limiting the molecular tools available.</p><p>Methodology/Principal Findings</p><p>Recent advances in magetophoresis and microfluidics have been employed to produce an automated platform called LiquidBiopsy®. This platform uses high throughput sheath flow microfluidics for the positive selection of CTC populations. Furthermore the platform quantitatively isolates cells useful for molecular methods such as detection of mutations. CTC recovery was characterized and validated with an accuracy (<20% error) and a precision (CV<25%) down to at least 9 CTC/ml. Using anti-EpCAM antibodies as the capture agent, the platform recovers 78% of MCF7 cells within the linear range. Non specific recovery of background cells is independent of target cell density and averages 55 cells/mL. 10% purity can be achieved with as low as 6 CTCs/mL and better than 1% purity can be achieved with 1 CTC/mL.</p><p>Conclusions/Significance</p><p>The LiquidBiopsy platform is an automated validated platform that provides high throughput molecular access to the CTC population. It can be validated and integrated into the lab flow enabling CTC enumeration as well as recovery of consistently high purity samples for molecular analysis such as quantitative PCR and Next Generation Sequencing. This tool opens the way for clinically relevant genetic profiling of CTCs.</p></div

CTC Flow cell operation and performance.

<p>(A) The LiquidBiopsy CTC flow cell. (B) A cartoon illustrating how target cells are pulled from the sheath flow while non targets move through the flow cell unhindered. (C) Efficiency of recovery of target cells using EpCAM based recovery: Incremental numbers of MCF7 (N = 34), HCC1419 (N = 27) or A549 (N = 65) cells were spiked into NHD blood and purified on the CTC flow cell. Recovered cells were enumerated in the flow cell. Graphs show number of targets spiked per mL of blood against the yield +/−1 SD from an average of between 7 and 27 experiments.</p

Direct molecular detection of tumor cells recovered from blood.

<p>Results of a TaqMan PCR mutation assay for the EGFR Exon 19 deletion mutation in H1650 cells. DNA recovered from incremental numbers of H1650 tumor cells spiked into whole blood and processed on the LiquidBiopsy platform are indicated. The mutation is undetectable in whole blood even at the highest input.</p