Home Forums Intestine Ask Me Anything: Modeling IBD on the Colon Intestine-Chip

Ask Me Anything: Modeling IBD on the Colon Intestine-Chip

Home Forums Intestine Ask Me Anything: Modeling IBD on the Colon Intestine-Chip

    • Rebecca Bonfig

      On June 2, 2023, starting at 11am EDT, Marianne Kanellias and Chris Carman will be hosting a live AMA (Ask Me Anything) session to discuss how researchers can accelerate drug development for inflammatory bowel disease using Organ-Chips. Check out the on-demand webinar here:

      Accelerating Drug Development for Inflammatory Bowel Disease with Organ-Chips

       

      Feel free to submit your questions in this thread before or during the event. Click the subscribe button to follow along for live updates!

    • Lena Neufeld

      Hello,

      How do you confirm the cells in your chip behave as in the body? In more than just morphology..

      Can you use your chip for microbiome inclusion?

      Thank you,

      Lena

      • Marianne Kanellias

        Hi @Lena Neufeld, thanks for your question! We assess several complementary readouts from the Organ-chips to verify that the cells are behaving as closely to human in vivo as possible. Specifically, for the Colon Intestine-chip we observe:

        • Immunofluorescence staining to verify the presence of cytoskeletal and tight junction markers (ZO-1, E-cadherin, F-actin)
        • Apparent permeability assay demonstrating formation of tight barrier
        • Bulk RNA-seq analysis can be performed on cells extracted from the Organ-chips, demonstrating closer gene expression to in vivo than organoids
        • Multiplex effluent analysis can assess soluble factors excreted from the vascular or epithelial channels

        I would also check out this Colon Intestine-Chip publication for more information: https://www.cmghjournal.org/article/S2352-345X(21)00145-4/fulltext

        To answer your question on microbiome inclusion, yes it is possible. Here is a study published from the Wyss Institute using Organ-chips to study anaerobic bacteria on intestine: https://www.nature.com/articles/s41551-019-0397-0

        Let me know if I answered your full question!

      • Christopher Carman

        The cells in the chip have all been extensively characterized in terms of function (e.g., barrier properties, mucin production), morphology (e.g., mature brush boarder with well differentiated microvilli and tight junctions) , multi-lineage epithelial cell subtype formation at expected ratios and transcriptome. All of these properties closely align with what is observed in human tissue. Much of the characterization can be found in our following publication: <span lang=”EN-US” xml:lang=”EN-US” data-scheme-color=”@F4F4F4,2,18:20000,19:80000″ data-usefontface=”true” data-contrast=”none”>Apostolou A</span><span lang=”EN-US” xml:lang=”EN-US” data-scheme-color=”@F4F4F4,2,18:20000,19:80000″ data-usefontface=”true” data-contrast=”none”>, et.al, Cell Mol Gastroenterol Hepatol. 2021;12(5):1719-41.</span>

      • Christopher Carman
    • Ibrahim Erbay

      How did you select the flow rate for your device? How does it relate to the colonic flow?

      Thank you

      • Christopher Carman

        The flow rate of 1000 ul/hr is the maximum flow rate that is achievable in our current instrumentation. It provided a fluid shear force of 0.3 dyne/cm2, which is a the low end of the physiologic shear forces seen in the post-capillary venule of the intestinal microvasculature , where most immune cell traffic and the normal shear rates are ~0.5 – 2 dyne/cm2.

         

    • Ben Gossart

      Hi Chris and Marianne,

      Thank you for an awesome webinar! I’m curious if any IBD therapeutics besides anti-TNF therapies have been tested on the chip. Further, what does this workflow look like in testing new therapeutics?

      Thanks!

      • Christopher Carman

        Yes, indeed  we have evaluated a number of other clinically relevant IBD therapeutics on this system, including dexamethasone (a classic corticosteroid), Entyvio (a biologic that blocks immune cell adhesion to the endothelium) and AJM300 (a small molecule inhibitor of cell adhesion). Our early goals were to validate diverse drug types across a multitude of mechanisms of action. We have shown that all of the above show efficacy in our model with the expected MOA and that we can rank order them with respect to each other.

        In addition, to selecting validated clinically relevant drugs, we also align dosing with clinically relevant Cmax (maximum blood concentration in patients) to ensure human relevant concentrations are being studied. With all of this in mind we have built confidence that this approach will be valuable for evaluating new therapeutics that our customers are developing in their IBD pipelines. In deed for the past year, we have been working with a multitude of such customers and seeing promising results.

    • Jackson

      Hello Marianne and Chris,

      Do you have all the cells in the system? Enterocytes, Goblet Cells, Enteroendocrine cells, Paneth Cells

      Thank you

      • Christopher Carman

        Yes, indeed, our model captures all of these cell types and importantly, they are present at the expected relative ratios to each other. The model also includes stem cell population that aligns with what is seen in vivo that helps to maintain the above populations.

         

      • Marianne Kanellias

        Hi Jackson, thanks for your question. As Chris mentioned we see that the Colon Intestine-Chip supports heterogeneous cell types present in the intestine: absorptive enterocytes (Villin+), goblet cells (Muc2+), enteroendocrine cells (ChgA+), cycling stem cells (Lgr5+) and quiescent stem cells (Bmi1).

        This publication assessed the percentage of these populations for three epithelial organoid donors: https://www.cmghjournal.org/article/S2352-345X(21)00145-4/fulltext

    • Rebecca Bonfig

      Hi Marianne and Chris,

      Really interesting work! I’m curious about the buoyancy media that you used in this model. What was your criteria for determining the composition? Did you notice any negative effects on the cells?

      • Marianne Kanellias

        Hi Rebecca, thanks for your question. The buoyancy media composition was developed to replace the presence of red blood cells in the vasculature which provide buoyancy, pushing immune cells to the blood vessel walls to roll along the endothelial surface. We use Percoll (colloidal silica) to create this buoyancy in the channel during flow so that the immune cells interact with the membrane. We also add gellan gum (polysaccharide) to homogeneously suspend the immune cells when not under flow (i.e. in the Pod media reservoir).

        This buoyancy media solution is based on standard RPMI-1640 media and provides necessary nutrients for immune cell culture. We do not see any effect in immune cell viability or activation through flow cytometry analysis of immune cells in buoyancy media up to 72hrs, including after flow through the chip.

    • Marianne Kanellias

      Thank you everyone, from Chris and I, for joining today’s session! This concludes the live segment, but feel free to keep posting any additional questions in this thread and we’ll try to answer them.

    • Anthony Heng

      Hi Marianne and Chris,

      Have you looked into assessing variability in PBMC recruitment and barrier disruption between different PBMC donors or different intestinal organoid donors? Additionally, have you seen any differences in effects of IBD therapies (i.e. Entyvio, AJM3000, etc.) between different donors (either PBMC or instainal organoid donors)? Thank you!

      • Marianne Kanellias

        Hi, thank you for your question. Firstly as a note on how we select PBMC donors: we select for non-inflammatory conditions (Non-smoker, non-obese, etc.) as well as perform immunophenotyping to ensure all immune cell populations are within standard healthy range.

        We do see expected variation between multiple PBMC donors in the magnitude of  recruitment response, as well as in downstream barrier damage. Currently we are expanding our immunophenotyping to assess responsiveness to our cytokine treatment. We also see a variation in therapeutic response between PBMC donors for small molecule, corticosteroid and biologic drugs tested. This may be aligned with clinical data since IBD therapeutics have historically lower response rates from 50-60%, and even lower remission rates ([1], [2]). We have yet to determine which factors indicate a responsive PBMC donor – this is still a burning question for predicting patient response clinically.

        The work so far has not investigated multiple intestinal organoid donors, but it would be a useful avenue to pursue to build robustness.

        Hope this helps address your question

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