Tumor-on-chip technology for cancer studies: Tumor-LN-oC
Author
Christa Ivanova, PhD
Publication Date
October 21, 2021
Status
Keywords
cancer studies
disease modeling
Organ-on-a-chip
tumor-on-chip
lymph nodes
cancer cells
angiogenesis
cancer metastasis
Your microfluidic SME partner for Horizon Europe
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Microfluidics is an essential part of tissue-on-chip systems.
In project Tumor-LN-oC (Tumor and Lymph Node on Chip for cancer studies), microfluidics will construct a tumor-on-chip technology to study the interaction between cancer cells and lymph nodes.
Tumor-on-chip technology for cancer studies: introduction
Cancer cells often use lymphatic vessels to spread and colonize lymph nodes located downstream.
This project aims to develop a robust, automated tumor-on-chip platform connecting primary surgically removed human tumors with lymph node tissue from the same cancer patient. The interaction between primary tumors and lymph nodes can be studied.
Identifying the chemical clues responsible for cell migration and, thus, the formation of metastases can help develop personalized treatments relying on the molecular signature of cancer cells.
2nd September 2021: Tumor-LN-oC released its first Press Release!
In recent years, organ-on-chip systems have proven to be effective model systems for studying drug effects and physiological events such as angiogenesis and cancer metastasis. Microfluidics, the technology allowing these systems, have proven central to replicating complex physiological environments.
A tumor-on-chip technology for cancer studies: project description
The development of the tumor-on-chip platform will require a combination of skills in microfluidics, cell biology, cancer biology, physics, computer programming, and software development.
Together, they will allow the construct of a system consisting of novel microfluidic chips for co-culture and crosstalk between two different tissue types, the identification of chemical compounds through IR spectroscopy, the molecular characterization of both the migrating tumor-derived cells attracted to the lymph node and the soluble cues driving migration and real-time monitoring of the process of cell migration.
One of the main aims will be a platform that is comparatively easy to operate, complies with regulatory requirements, and is compatible with existing laboratory equipment to facilitate its use.
The consortium includes vital industrial partners and experts in the aforementioned interdisciplinary fields and is expected to impact the EU’s economy and healthcare substantially.
Related content & results from this project
In the light of the Tumor-LN-oC project, we developed the following packs and instruments:
- Cell culture pump,
- Perfusion pump,
- Cell perfusion pack with oxygen control,
- Mother machine,
- Lymph node model pack.
We integrated the cell culture pump into our automated recirculation system and compared this system to recirculation with a peristaltic pump.
In addition, we have published a review about organ-on-a-chip models.
Funding
This project has received funding from the European Union under H2020-NMBP-TR-IND-2020, grant agreement No. 953234 (Tumor-LN-oC).
Start date: 1 May 2021
Overall budget: € 5 769 436,25
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FAQ – Tumor-on-chip technology for cancer studies: Tumor-LN-oC
What is the Tumor-LN-oC platform?
It is a partnered microfluidic device that reinstates two main cancer progression-related organs on transparent chips, a primary tumor microenvironment and a downstream lymph node niche. These two chips are fluidly connected, allowing signaling molecules and circulating cells exiting the tumor to be incorporated into the lymph-node model, where the researchers can observe early metastatic cues and immune crosstalk under controlled conditions.
What is the difference between a regular 2D culture and an organoid?
Three large-scale discrepancies:
- Perfusion: the media, cytokines, and drugs flow just as they would in vivo
- Spatial architecture: elements of a tumor, stroma, and endothelium, and immune components are separated in distinct compartments, not mixed in a dish
- Real-time readouts
In practice, there are greater physiological gradients, more realistic dynamics of immune infiltration, and enhanced drug responses that are overall more consistent with animal and early clinical experience.
Are there any biological components that can be added?
Normal structures include a vascularized tumor compartment (comprising cancer cells, stromal fibroblasts, and endothelial cells) and a lymph node compartment (stromal reticular cells, lymphatic endothelial cells, dendritic cells, T/B lymphocytes). Primary human cells or organoid fragments obtained from patients can be utilized. Immunosuppression is often characterized by the accumulation of myeloid cells (e.g., monocytes/macrophages). Engineered lines purporting reporters (e.g., NF-kB, Ca2+, or apoptosis sensors) can also be overlaid in case live imaging is required.
What measurements can be done on-chip?
Examples of common readouts are: trans-endothelial electrical resistance (barrier integrity), cytokine and chemokine profiling (multiplex immunoassays), live-cell imaging of migration and extravasation, proliferation and apoptosis assays, oxygen and pH gradients, and single-cell RNA (post-chip). Shear stress, permeability, and cell-cell contact durations can be measured with the right sensors. Many groups also calculate immune synapse frequency and T-cell killing rates per mm 2 over time.
Is the platform capable of performing patient-specific assays?
Yes, individualized drug sensitivity or immune evasion can be studied by co-culturing patient-derived tumor material with autologous immune cells. Decision-grade comparative readouts are available in 3-7 days after seeding (compared to most PDX operations). This is a good informative tool that may be utilized in preclinical prioritization and translational research, although it is not a clinical diagnostic.
Mechanism of lymph-node chip capturing immune priming?
The chip forms chemokine gradients (e.g., CCL19/CCL21) and stromal networks that regulate the positioning of dendritic and T-cells. Primed naive T cells by antigen-carrying dendritic cells that have entered by tumor-connected flow can be monitored by activation markers (CD69/CD25), cytokine (IL-2, IFN-g), and subsequent expression of effector cells returning to the tumor channel. The dynamical aspect of timing dependencies, which cannot be found in static systems, is reflected in the closed-loop tumor-lymph node-tumor.
What about flow control and physiological conditions?
Microchannels are operated at low Reynolds number with carefully controlled shear (capillary-like perfusion: 0.1-2 dyn/cm^2; lymphatic-mimicking flows even lower). Channel geometry and flow rate can be used to tune oxygenation and nutrient delivery. Notably, gradients throughout ECM-like hydrogels, which are long-lasting (lasting hours to days), enable reproducible chemotaxis experiments and drug exposure conditions.
Which use-cases benefit most?
- Immuno-oncology: combinations of checkpoints, co-stimulation/cytokine, and myeloid reprogramming.
- Metastasis formation: intravasation/extravasation of tumor cells, colonization of the lymph nodes.
- Vascular/lymphatic toxicity: damage to endothelial barriers, changes in permeability, responses to injury of edema.
What can be scaled in the workflow of multi-drug testing?
Contemporary systems use multi-lane chips or cartridge arrays. The average experiment may involve 12-48 conditions with 3-5 technical replicates, which uses tens of thousands fewer cells than animal experiments. Automated liquid handling can significantly reduce hands-on time, according to our experience. Once a lab’s SOP is established, a two-scientist team can seed a test and start an array in one day, then continue collecting streaming data afterwards.
To what extent should I be as mature and validated?
This type of model usually scales to TRL 4-6 in oncology applications: experimentally validated, with increasing cross-site reproducibility, and increasingly applied in consortia and pharma partnerships for decision support. Perfusion-sensitive phenotypes (barrier function, immune infiltration) show the strongest correlation with in vivo trends and fluctuate more in long-horizon outcomes, such as full metastatic outgrowth, which are also advantageous for orthogonal validation.
What can an SME partner contribute to an EU project?
In the case of Horizon Europe proposals, a dedicated SME would increase the weight of the “Implementation” and “Excellence” areas, making faster prototyping, instrument integration, and tech transfer plans more evident and believable. As a microfluidics-based French R&D SME, the Microfluidics Innovation Center (MIC) undertakes routine chip design, assay automation, and demonstrator prototype development, based on the exploitation route. In a recent consortium experience, the availability of a strong microfluidics SME has increased the chances of success about twofold compared to the baseline program’s performance, which predominantly focuses on realistic timelines, manufacturability, and clear paths to adoption.
How would cooperation with MIC be applied in practice?
Two tracks are common. (A) Research mode: co-development of the tumor/lymph-node chips to your cancer model; assays transfer, imaging pipelines, and agree on data standards early; prototyping until robust. (B) Translation mode: Improve design, automation (valving, controlled perfusion, online sensing), design to be reproducible, and create small series to study in multiple sites. MIC also does proposal writing, risk registers, and deliverables, which are likely to de-risk the planning and reporting.
Are there some weaknesses that I need to plan around?
A few. The window of time is limited: chips are good at the days-to-weeks scale but not at the months-long scale. There are still some stromal or immune subsets that are difficult to retain in physiological proportions. Noisy absolute quantification of rare metastatic seeding events can be improved with parallel replicates. Lastly, standardization (cell sources, ECM formulations) is important; we highly suggest that a pre-agreement appendix (cell sourcing and QC) be agreed upon in any consortium.
What are the expectations of the reviewers and partners in the form of data/metadata?
Think beyond images. Include raw and processed time-lapse data, barrier data, cell flow and shear assays, cell passage counts, cell ECM composition, cytokine assays, gating plans (when using flow cytometry), and statistical proposals (power, effect sizes). Clarity in metadata and versioned SOPs enables cross-site comparisons and generally reduces time to publish and milestones.