Advancing Discovery by Enabling the Use of 3D Organoids

This talk was presented as a part of the Bioprocess Quality, Data and Analytical Solutions webinar with BIO-EXPO. Hear from Victoria Marsh Durban, PhD, Director of Custom Organoid Services at Molecular Devices about how Molecular Devices is overcoming the unique challenges that 3D in-vitro models bring using innovative technologies to enable high-throughput robust screens using powerful cell models. Victoria discusses how organoids are used in the drug discovery process and how these models can carry through to the full preclinical development phase.

Victoria takes the audience through:

The challenges with bringing a drug to market
Current models used in drug discovery
What organoids are
Challenges with 3D organoid models
Solutions from Molecular Devices
A case study
How to access 3D ReadyTM Organoids

Transcript

Our first three presentations are brought from three different brands, all from Danaher. They are Molecular Devices, Beckman Coulter Life Sciences, and SCIEX. First up is going to be Victoria Marsh Durbin. She is the director of custom organoid services.

She is going to present ways in which molecular devices is overcoming the unique set of challenges 3D and vitro models bring using innovative technologies to enable high throughput robust screens using these powerful cell models. However, first some words from Thomas Colunga of Danaher.

Thank you, Perry, and thank you for the introduction and welcome to everyone here today. I just want to take a brief minute to introduce Danaher and who we are. Danaher is a leading science and technology company as evidenced by our number of leading life sciences brands here on the screen. We have a mission and a vision to help reduce the time the market of life saving therapies by 50%. And the way we plan on achieving that is becoming a critical partner along your IND, INDA, and VLA path. So our commitment to human health is essentially to leverage the power of our science and technology companies and engineer high impact solutions that have an impact on improving health outcomes for our patients and patients around the world. We leverage that by leveraging our people, our capital and our DBS, which is our Danaher business systems. And so what I'm showing here is just a brief snapshot of the companies in our portfolio. A few of the companies calling out specifically three of them that you're going to hear from today.

Molecular devices, Beckman Coulter Life Sciences, and SCIEX. And so I thank you for joining us today and I hope that you find our presentations very insightful. So without further ado, what I'd like to do is pass things over to Victoria.

Great. Thanks so much, Thomas. And good morning. Good afternoon to everybody. Thanks so much for joining us today. And it's really great to be here to kick off this session today.

We're starting off with a very quick poll question here. And while you think about that, I'm just going to talk a little bit about what I'm going to be speaking about today. So today I'm going to be presenting on the use of organoids within the drug discovery process. Now, I appreciate that I'm going to be moving a little bit upstream of typical CMC, which I think is where the focus of this whole event has been. So really thinking a little bit more about how we use models within the drug discovery process, and then of course how that can carry through to the full pre-clinical development phase.

So really, I don't think this slide needs any introduction to this audience. I think it's a problem that we are all familiar with, which is that drug discovery today is costly, it takes a lot of time, and an awful lot of resource. And over time, this is a problem that is not improving. Now, molecular devices, we believe that part of the reason for this problem is that human relevant models are not being used early enough within drug discovery. So here's a range of different models that people tend to use within drug discovery. So it ranges all the way from simple 2D cell culture models, all the way through to much more complex organoid-chip type systems. The current paradigm is that the models that are used today are primarily 2D cell cultures, which are a very simplistic model and have the advantages of being very easy to use and very inexpensive.

However, these model systems don't always give very representative data, and therefore don't translate to how a patient might respond to a candidate drug therapy. So just in terms of what organoids actually are, I appreciate, you know, people may have heard of organoids or may or may not understand the terminology or may think that organoids are something different. But the definition that we work by is that the minimum definition of an organoid is that it contains more than one cell and more than one cell type. Organoids grow in three-dimensional space, and they self-organize themselves spontaneously to more faithfully recapitulate the tissue structure and critically the function of the tissue from which they originate. They can be considered as simplified many organs or indeed many biopsies if they come from the disease state, and organoid growth is driven by stem cells. And this has the advantage of the fact that it allows propagation of organoids and culture over multiple passages to be able to produce a lot of material to perform your experiments.

So in the graph that's on the center of this image, really what this is showing is the current paradigm is to use two-dimensional cell lines. As I already mentioned, the benefit of this is that they're highly scalable and relatively inexpensive. However, the payoff for that is that their translatability is not particularly high. On the opposite end of the spectrum, we see animal models which have good translatability.

However, they're not scalable, relatively expensive, and obviously ethically questionable in terms of their widespread use. So what we at Molecular Devices believe as a new paradigm for drug discovery is to use three-dimensional in vitro models. As I've already said, these are more representative and predictive of human response and can replace animal models. And indeed publications have shown that these 3D models can shorten the preclinical development time, which leads us to the hypothesis and to this proof statement, which is that we believe that using 3D human relevant models can accelerate drug discovery to be able to identify successful drug targets twice as fast as using the current paradigm. Now, of course, with the use of more complex models comes challenge. And that challenge is brought about by the fact that these models require greater expertise and more skill to use. There are a few robust and reproducible protocols to be able to use these. They're relatively difficult to scale compared to simpler systems.

There can be variability within culture of material. They're slated to produce. They're difficult to automate. And in general, there's high cost and investment needed for the use of these models. And these are really the challenges that we're looking to address at Molecular Devices to enable the use of these models more widespread and more different applications than they're currently used. So just taking a step back then to look at the broader Molecular Devices portfolio, I think most people are very familiar with the fact that Molecular Devices is really known actually in the microplate reader business and more on the analytical side and also particularly in the drug discovery space for our flipper instruments. However, we also have a footprint in a number of different areas. And one area that has been rapidly developing within Molecular Devices is the three-dimensional biology space.

And within that space, we have these four main areas. So, and these are focused on hardware and instrumentation for automation of cell culture through our Cell Express AI, which is our newly launched, fully automated in a box system for the culture of 3D, Organoid and spheroid models. We also have our Bioassembly Bot 3D Bioprinter. We also have our Organoid Innovation Center, which is focused on developing custom automated workflows for the culture of three-dimensional models and the screening of those. And where I'm going to focus today really is on our Organoid Expansion technology, which is relatively new addition to our portfolio, which is technology for the production of organoids at scale.

I'm really focusing on the industrialization of these complex and highly powerful models. So, some of the advantages of our Organoid Expansion technology is that we manufacture the organoids at scale here on site, where I'm sitting today, which is in Cardiff in the United Kingdom. We produce the organoids at scale using our proprietary bioactive-based bioprocess. And this has all the advantages that you would expect of using a bioreactive-based bioprocess to produce a product at scale. So, we can produce organoids at high scales.

So, at the moment, up to six million organoids in a single batch. We have great reproducibility, both batch to batch and within a batch. The product comes with full traceability and quality assurance.

So, again, looking at really leading the industry in terms of the QC and QA type attributes, we're putting on this product to enable the adoption of these models into a more regulated environment. The organoids that we produce are very simple to use. They come as a ready and frozen format and they can be used straight out of the vial. So it's basically plate them, allow them to recover and then go ahead with your assay. So the turnkey end to end solution for getting data is sort of a seven to ten day window.

So very fast and very simple. One question that we typically get asked by customers is, you know, the people tend to get a bit scared by a bioreactor process, you know, particularly for these sensitive and complex models that could be impacted by putting them through such an industrialized process. So one thing that we've taken some time to do is to really reassure our customers that actually the process that we put the OrgNoise 3 for expansion doesn't actually impact the biology of the actual organoids themselves. So in this particular case study that I'm presenting here, we set out to compare your standard manual culture expansion with all manual steps and no freezing step and the analytics of that to our bioreactor based expansion, whereby we produce our organoids in 3D ready format.

We freeze them down and then we throw them just before assay. And we subjected these OrgNoise that we're using here, which in this case actually is some breast cancer organoids to a range of different analytical methods so that we could compare the manually grown organoids to the bioreactor grown organoids. And I guess without too much of a spoiler alert, what I'm going to tell you is that we saw very little difference in terms of how the organoids performed in assays when they were grown either through the manual process or through our bioreactor process.

So this just gives you a bit of an example of the organoid line that we were using. So it was a breast cancer line which expressed some characteristic markers of breast cancer, which we saw these markers were unchanged both pre and post bioreactor expansion. We looked at the driver mutations, the genetic background of the organoid line to see how that was changed between manual and bioprocess expansion and we saw absolutely no differences. Then going again to a slightly more sensitive assay, which is looking at gene expression analysis. Again, we were unable to detect any significant changes between organoids that were expanded through the bioreactor process and phrasing and thought immediately prior to the assay versus those taken from manual culture and straight into assay. So again, very few changes and indeed very difficult to discern the differences between lines that were cultured manually versus those in the bioreactor.

And very finally, we looked at some typical standard of care drug responses in this organoid line through standard drug assays. So these were performed manually rather than through an automated or a liquid handling solution. These were just very small scale manual assays very much to recapitulate what customers might do in their own hands in the laboratory.

And again, what we found is that the drug response and the error bars, so the reproducibility of these assays was improved in our bioprocess grown organoids versus manual culture. So overall, what we're saying is that we have the advantage of scalability without actually impacting on the model itself. In terms of how our customers can access organoid expansion technology, there are really two main ways to do this. The first is through being able to purchase our 3D ready organoid vials off the shelf. So if you're not sure that organoids are right for you or you want to explore the technology, see how easy it is to deploy into your existing assays.

You can come to us to purchase its single vials of organoids off the shelf that we can provide to you as a consumable. Alternatively, if you already have an organoid line that you're working with and you know that you want to work in this model system, we provide a completely custom and bespoke organoid expansion service whereby we will work with you to be able to expand your organoid line, develop QC package and QC and QA testing completely bespoke for you. So again, really just to hammer home the benefit of working with the 3D ready organoid products is really this time saving. So compared to manually producing organoids where you need to throw your organoids go through multiple cycles of expansion before you have enough organoids to go into your assay, with the 3D ready organoid products you can take organoids from the freezer all produced in the same lot. So you could have done your qualification and your validation of the product all in one go for that particular batch.

When you're ready to run your assay, you take the vial from the freezer, throw it and plate it, allow it to recover, go ahead with your assay and you've basically got the data that you need within 7 to 10 days compared to needing to do 30 days or more if you're doing multiple rounds of manual culture. So again, just to come back to this slide in terms of what I've said today. So I already mentioned that molecular devices is on a mission to really reduce some of these hurdles to adopting 3-dimensional organoid organoid culture and organoid technology and some of these I've addressed today. So the ones that I've grayed out here are the ones that I've spoken about today that we overcome with our organoid expansion technology. The other ones on this slide have actually been addressed within other areas of molecular devices through the automation and the development of automated protocols for organoid culture and organoid assay. And so, you know, what I'm trying to say here is that I haven't been able to tell you everything today about what we've got going on in the molecular devices portfolio in 3D biology but we are addressing all of these areas of challenge in one way or another. And with that, I will finish and thank you for your attention. I'm very happy to take questions and please contact me afterwards if you're interested in finding out more about organoid and 3D biology solutions.

Wonderful. Thank you, Victoria. We have a couple minutes for questions. So, which steps of the organoid culture process can be automated and can you also speak to some of the advantages of automating? Sure.

So actually almost every stage of the organoid expansion process can be automated. So we have this already happening within our labs within molecular devices. Our application science team are highly talented, very skilled at moving manual processes into automation processes. Really, yes, we can automate pretty much every step. So whether that's, you know, the thought and the initial plating, the breaking of matriarchal domes, the breaking of any other hydrogel domes, the breaking of organoids to be able to dissociate them and replate them to expand the cultures, the addition of compounds or other cell types or T cells, all of these steps can be automated. Of course, some of this is built into our Cell Express AI platform. So this can all be done in an automated manner. And what that does is also builds in automated imaging and decision making so that you can completely, in a completely unbiased approach, you can decide whether an organoid culture needs to be fed, needs to be passaged, is ready for assay indeed. So, you know, that's, again, going to the second part of the question there. That's one of the advantages is the removal of bias and the subjectivity in terms of, you know, do my organoids look good?

Does the culture look good? Is it ready to go? Is it not ready to go?

Does it need something else? Plus also, obviously, the benefits of automation is just that all the process steps are always done the same time, every time. So, you know, there's a lot of variability operator to operator, particularly with the very manual hands-on steps that are involved in organoid culture. So you might think about, you know, like how hard and fast you're pipetting, you know, exactly how fast you add or remove your media. So all of it, how fast you can do the whole process, because all of these individual steps actually have a significant impact on the reproducibility of organoid cultures. And obviously automation then will take out all of that variability, all of that operator variability, and also free up operators to go away and do things that are more valuable than your straightforward organoid culture.

Very good. How can I be sure that the reactor, the bioreactor process doesn't change the properties of my organoid line?

Yeah, so again, so this question I've covered a little bit already in terms of, you know, I've shown you the case study of how, you know, we've looked at that one particular line pre and post. You know, again, where we work on custom projects for customers, we can develop completely bespoke QC packages, whether that includes, you know, micro expression analysis, whether that includes, you know, genome-wide type RNA-seq or genetic analysis. We can really build that to give the customer that assurance that there's no change in the biology of the line.

Excellent. Well, thank you, Victoria. Thank you for being with us today. That was Victoria Marsh Durbin. She is with Molecular Devices, part of Dan and her.