 Cool, so good morning, my name's Deepak and I'm a research fellow with the synthetic biology center And so today we're just going to talk about kind of how do we apply synthetic biology for augmentation in the capacity of talking about organoids and so where I first want to start is About 25 years ago We saw actually an ear on the back of a mouse from Linda Griffith here at MIT and some other researchers in the Boston area And you start thinking to yourself well, wow, we're building tissues But in actuality it's just some polymer on the back of a mouse that you've squirted some cells on and so you've grown a structure Fast forward 30 years we can say well I can 3d print that same structure now in an hour which used to take five years to make just because you had to make the Polymers you had to understand how to implant them But really all we've done is build a structure That structure is just a great thing to put some earrings on but in reality. It's not functional By no means is this actually going to be a functional ear by any means So we have to start thinking about how do we actually grow and augment tissues in a way that actually gets past This structural specification and actually incorporates function and the tools of synthetic biology is actually something we can do We have some proteins and genes these protein and genes come together in biochemical reactions to form interactions Those interactions then become pathways that can now encode sensing processing actuation and in turn leads to a cell Groups of cells become tissues and cultures and all of a sudden we can start thinking about how do we actually engineer underlying? elements to create functional novel behavior So this is always a good time to pause and reflect that a cell robots biobots Whatever you want to call them often benefit from establishing kind of engineering paradigm So we can think about this a little bit more deeply if we had to engineer a cell from scratch We probably wouldn't put the mitochondria and the ribosomes exactly where they are But we can use kind of this sensing processing actuation and start brainstorming the fact that there are many modalities Accessible to us in this programming nature. I can go at the chemical molecule. I can use proteins. I can use mRNA I can use all of them all together and we actually put all of these together to start thinking about how do we build systems? This is where I usually take a pause and say While we often think about digital logic and this very ordered cell courtesy of Tim Lee where we can program anything It's always healthy to know as we are before lunch that a cell is closer to a burrito So we have to start thinking How do we harness biology? How do we augment these things? And how do we actually get to a place where we can build systems using biology? So let's take a case of I wanted to grow a new liver a Program that you might think about is if I have a single stem cell I'm going to write a genetic program that has it actually multiply So now I have a collection of IPSC cells I have to go through some symmetry breaking operation that yields now these prototypical cell types That can become this actual potential liver So rather than just putting some polymers down and squirting cells on it Can I get through the symmetry breaking to make the hepatocytes the vasculature their actual neuronal tissues and in turn? grow into these organoids on a chip or actually true functioning organs and that's actually where we are today is how do we go through this? process of writing genetic programs for Augmented organoids as well as future organs that don't even exist right now Why is it that our livers can't see in the dark? Maybe they should have rods and cones as well? So that's where we are and so now for the next kind of few minutes I'm going to talk a little bit about where we are in this process and what kind of we can do so far so over the last 30 years there's been immense research efforts in IPSC's and All sorts of condition, but one of the things we've realized is cells actually go through a self-fate kind of Decision tree that can be manipulated in many ways Originally we discovered that using small molecules or the Yamanaka factors You could take cells both forwards and backwards through the decision tree of differentiation And so one of the things we would want to do then is how do we make this autonomous? How do we make it so that way we can grow new organs using these synthetic networks? So that might look like this where we have just some IPSC's IPSC's then can become either ectodome, mesoderm or endoderm So again going down this hierarchy of differentiation that is actually recapitulating Just embryogenesis here from their certain cell types will actually differentiate into kind of the specialized lungs, liver, stomachs, and you just keep going down and down where you have all of these Cell types that you need to navigate a cell down these fates and avoiding going down the other fates using a genetic program This seems very simple because it just looks like it's a binary decision tree and reality It is actually not as nuanced as that But you can still we try to write this program saying we want a self-timed differentiation multi-step process where I have an input chemical that starts down this progression and I incorporate feedback and Indeed, that's what we've been starting to do is this is now work that was published a few years ago Patrick Guy and Mo Ibrahim both of whom are familiar in the MIT community as we've all been here 10 years now But these endoderm then start expressing these self-fate regulators and you actually go through the symmetry breaking where you can have a Fairly simple circuit Integrated into these IPSCs that in the presence of a small molecule docks that is FDA approved You can now start having the first symmetry breaking happen where during day zero to five you would have this break You remove that input and the actual circuit will continue operating Hopefully getting into that first symmetry break So here's some experiments that we actually did where we introduce a mixed cell population They're all IPSCs to start but one is fated to become actually this endoderm and the others actually going to become ectoderm so the endoderm is these blue cells and The red cells are this ectoderm after administering docks You'll see that those blue cells start turning green showing that they've actually gone through that first cell differentiation We can go a little bit further and after that critical symmetry break Eventually we can actually start staining for downstream things So that miso derm is actually going to become the vasculature and we can start staining for blood vessels in these prototypical tissues We can take that endoderm and they actually become hepatocytes Starting to make albumin and fibronectin that we can see expression of showing that it is truly a hepatocyte and in turn we can actually Make all sorts of different cell types are ready. So just in this very simple circuit We've been able to make the three main classes miso derm endoderm actor derm and through actually very various Kind of coaxing downstream of that. We've now generated around 15 different cell types And now the hard part is is we don't know how to build a liver like truly But we know that all the cells in the liver we can map and they are all of these cells that we've been able to make So we're now in this kind of feedback cycle of how do we engineer new programs to get the cells where we want to Is indeed it is these very local rules that go govern this global structural specification and that's where we truly are So with that we've been working on a lot of things Here's like an organoid that I made yesterday and it is just 50,000 cells with different circuits This one is going to try to have differential adhesion so that we we have like a sticky inner core and an outer core and that's Kind of like where we are we live in the future. They are kind of ugly at times Here's a huge organoid that I made a month ago and they're actually like as Big as your fists if you actually put them all together Which is kind of cool to think about is right now It's not that unlike if I just stretch it over a cartilage scaffold. I can put it on my hand and say I have an ear so with that all this work takes many many people as This is probably around ten years of work both by myself by all of my collaborators by the entire white slab as whole And I'm always happy to chat about anything related to ears burritos and symbio