BIMM134 Metastasis I EMT and metastatic cascade

[00:00:00]Okay, so we're going to begin metastasis today. And uh as I've said with angioenesis, we're going to see a lot of the processes are going to be pretty similar as far as metastasis and angioenesis, including some of the same players like MMPPS and u and changing vinegar, things like that. So we'll sort of see it again some factoids uh that I probably have mentioned before too. Again, 70% of patients about have already metastasized prior to detection of cancer. And that is uh the most likely cause of death is metastasis, which is of course why we need to get much better at diagnostics that are much are detection and diagnostics that are are much earlier.

[00:00:46]Um and overall though you know the interesting thing about it is by the time things have metastasized you know you have millions of different cells entering the bloodstream at any given time but it's not that easy to do.

[00:01:00]So the idea is overall a very small percentage you know of these cells actually form what would end up being fully metastatic lesions and full macromeats. Uh so it's an inefficient process but you know when you're you're putting that many many cells in the in the uh in the body it does basically come into play. Uh just put this slide up here to show in a sense of how how potentially as we get more and more better at detection too we're going to be able to detect very small micromets and also even circulating cells as well.

[00:01:35]So detection should get better at this point. And uh as I've said, I'm not going to show the video with this, but these are some good videos here. The uh you know, both of these hallmarks, and metastasis, very similar. And again, looking at this concept that really emerged uh with the immune system and and with uh androgenesis is this idea of this tumor micro environment. the idea of tumor controlling sort of the cells and the matrix surrounding it, the tumor stroma to do what it wants. But again, we're involving the same thing, proliferation, migration of cells, and again using proteasis to to remodel. And so let's let's make these pictures a little bit bigger.

[00:02:20]So I have sort of course I still have trouble with this overall on this. So I have a couple of just different overall pictures that of the metastatic cascade kind of going through these steps. These pictures are sort of just an introduction to kind of again sit back and see a visual of the complexity of the system. Uh you know noting a couple important little points as you go. So again remember benign tumor is this idea where is not invaded underlying tissue but once it starts an endling uh you know invading underlying tissue comes to the point of again having these cells degrade this matrix on the way to the blood vessel we're going to be calling this introverbation in a minute entering the blood vessel and again you could see that very few cells actually will survive this journey but some do and then you have the opposite okay you have to have these adhere to a capillary bed.

[00:03:16]You know, sort of a rest in some sort of target organ and then it's got to end leave the uh the bloodstream that's extravate and do kind of the same process of degrading the matrix, migrating through and then establishing uh a metastasis.

[00:03:33]And this last step is going to be sort of an important step because just because the cells have made it to this journey doesn't mean that this new environment uh that it's in the new place it's trying to get to will actually be able to sustain its growth because again to get to this point they've created this entire you know tumor stroma which has been activated.

[00:03:56]You know this may not be at this case.

[00:03:58]So that might be a way that we attack this. Again, uh this this one kind of emphas emphasizes that too. Again, these are just you know pictures we're going to use to sort of uh to go through each step. But you have kind of this idea of proliferation angioenesis kind of those first five hallmarks. But then we're going to go to the idea of how these cells will sort of lose adhesion between one another because you're not going to have the entire marble kind of go through. And then again this idea of you know introversation into the bloodstream. We're going to talk about how they surround themselves to sort of survive it and then the idea of it resting into some target and then leaving the bloodstream exterization. But again this is the idea of this micro environment.

[00:04:47]In order for this to be a full-fledged, you know, killer met, it has to have the right environment, which is the whole second lecture, but literally be on on that aspect of it.

[00:04:59]And one more picture just again I like it just sort of visualizes it in one one phase. Looking again at this, you know, sort of uh getting through the initial again these are, you know, carcinoma type tumors.

[00:05:14]They have to kind of get through this layer here, enter the bloodstream. We're going to talk about it surrounding themsel with platelets for protection and then again reversing the process here. Seeing a very sim some of the similar molecules are going to be used here like proteasis will be used here as well.

[00:05:35]Okay. So here's an important sort of concept that has emerged which is sort of sort of interesting.

[00:05:45]We'll probably get this down so I can maybe start to to draw a little aspect of this. We have some shadow here.

[00:05:56]If I can minimize the shadow.

[00:06:11]Sorry about Well, we'll try to live with the shadow right now.

[00:06:23]Won't be too I should get this a little easier.

[00:06:31]Yeah, it won't be too bad when I do it.

[00:06:32]So, figure that out later. Anyway, so we're going to this idea of this, you know, sort of overall change of these cell types again for these, you know, epithelotype tumors, right? Not the connective tissue one, but the carcinomomas which have this epithelial structure to it. And the idea of this is, and this is again the carcinoma. The idea is epithelotype tissues are connected by certain connections and literally are supposed not to move.

[00:06:59]They're no, they're non-motile at this point. But in order for it to actually, you know, travel and as it goes through, you know, the matrix and all the way across, uh, they're gonna they're going to resemble meenl type cells.

[00:07:13]Again, membal type cells are the ones like fiberblasts and things like that which actually have a different shape and are able to actually be more modile.

[00:07:22]And so you sort of have this idea of in the primary tumor, they're having these epithelial. It goes through this transition of being meenl like and having mel like structure in order again to to go through this environment and then you know what's really interesting too is then there's a recon conversion.

[00:07:42]So this idea of the zenimo back to epithelial uh which now will have some of the characteristics cytokeleton inrins etc that will match its beginning. So it's literally making a transition during its journey. Then it's reestablishes its original structure which is sort of amazing.

[00:08:04]And again just different pictures showing the same thing. Some of them show you know better than others. uh but again you have this this concept again carcinoma and situ or issue going again you could see even cartoon wise they have a given structure but then you know when they get into uh you know this mezeno mode so that's EMT now they're able to produce proteasis to migrate they have a different shape and that will continue until they again form you know the the the secondary met and then again Here is hope they got on this thing too. Here's this idea of you know in order for this to become a micromet to a real full-fledged metastasis it has to then redevelop this reactive stroma. Okay. So we really sort of care about this concept of you know what have to happen to get to this stage that we've talked about again modifying the tumor micro environment to its advantage with these other cells. Well, this new one, in order for this to actually occur, you're going to have to have a redevelopment of this uh activated tumor stroma, which again, as we go through this, to me, that is a uh a a point to to to basically be aware of that even if you don't necessarily, you know, stop cancer or detect it early enough this if you can make it where it can no longer be a reactive stroma. And in fact, if we can then modulate these components to make sure that they keep the cancer at bay, uh, you know, that might be one of the most effective actual treatments.

[00:09:45]And so, let's make this even bigger cuz this is just a cool cool picture.

[00:09:52]And when we go through the steps, you're going to actually see again this idea of, you know, sort of this one process of occurring here that we're going to see that when we look at the primary tumor, okay, the epithelial type tumor, it is surrounded by a basement membrane. Okay, it's kind of a specialized extracellular matrix that this these cells are attached to just like nor epithelial cells are. Well, one of the first stages then of metastasis will be again the breakdown of this basement membrane and you can see sort of this the structure here in green is taken away that's part of this EMT but then once it gets in this case it basically formed a liver metastasis it reestablishes its epithelial uh basically type so it's a dynamic process the cell takes uh the ones which can become metastatic here uh actually didn't transform and then go back to this because this is how they originally developed and this is how they develop that that stroma.

[00:11:02]Okay, a couple other aspects of this I'll probably put this on that wants to know about this EMT ha has to do with the connections between between the cells and so again if you're looking at when we're looking at normal epithelial type they're connected by uh by molecules called eadherin okay so one is the switches to EMT is going to be the loss of ecadherin and we'll see that also involves cell signaling the loss of ecadhirin and more of a uh a cell attachment called encadherin which is more what's on these messenal type. So that's part of EMT and in fact let's actually start sort of writing for EMT.

[00:12:01]Oops. That's why I know EMT sort of changes will sort of be loss of EAN and we'll show how that's important as well.

[00:12:31]and sort of then corresponding gain in basically shown there and cadherent attachment between cells.

[00:12:49]Okay. So again, here's a bunch of other sort of u sort of changes that are going from basically epithelial to memo. Again, the only ones I care about or are sort of pointing out is sort of ones which are important.

[00:13:05]Again I already put n cadherent but again basically to make it sort of simple increases I'm not going to have us remember is the specific ones but increases the changes in cytokeleton cytokeleton that basically makes it more fibroblast like without going to the specifics of it for this thing. Okay. The other ones too is the idea of because of this fiber like they do secrete fibonnectin and this is extracellular matrix which is important for when we start talking about inrines. Again inrins again bind to specific matrix components. Well, for meal cells like let's go ahead and kind of emphasize for the purple here.

[00:14:11]Uh they one of the main binding uh extracellular matrix components is fibonnectin. So we have that and also which obviously is important for we kind of realize it. Don't worry about the specifics, but it's also then going to secrete these MMPs.

[00:14:37]Okay. So there's other losses obviously associated with the loss of the epithelial uh phenotype but sort of concentrating then except for the loss of eadherin which will have some special importance will be the gain of these the gain of the different cell cell these are all again cell cell connections uh changes in cytokeleton that allow it to be you know to increase motility and also part of that is what extracellular matrix that they're secretreting but of course also MMPPS because again they are going to have to degrade the matrix and so the way I've kind of done this and you'll see sort of uh even the summary slide at the end is as far as uh you know using this type of figure at least to begin with you know we we kind of mentioned that a couple minutes ago too again the you know ankcoenic and tumor suppressor type mutations kind of get us through most of those first fall, you know, five hallmarks we talked about, uh, including androgenesis and kind of leaves us with that sixth one, which is going to be, uh, you know, basically metastasis.

[00:15:50]And so I'm going to go ahead and erase this and start to draw out some details of what we're going to call the metastatic cascade. sort of again six or seven steps that are involved in uh in this idea of metastasis. So we'll get rid of this. You sort of have it.

[00:16:21]So eventually I'll try to get rid of it.

[00:16:26]And so I'll be adding some details with this a bit and then we'll see kind of a summary slide that I also will will have up here.

[00:16:41]It says okay. So we roll to put over here.

[00:17:04]So, metastatic cascade and again we'll go ahead and almost repeat this but we're going to add some details of this. So again we have this idea the cells have to detach from one another and we'll see what this does. So again basically we'll put cell detachment.

[00:17:31]I'll show some pictures of it.

[00:17:36]And again one of the main things is again as I've already mentioned is the loss of the cell subconnection loss of e could hear it.

[00:17:59]And so what I like about should I make this a little easier.

[00:18:15]I like about this cartoon version of this is you really start to see then this idea and I don't know if I have room to put over here too of again the cells have to detach from one another.

[00:18:26]One of the main connections between uh epithelial cells is this ekadherin.

[00:18:32]Okay, eadher on both sides. We'll talk about what that means, but also we'll go ahead and kind of show here too. One of the next steps will be the loss of this connection between these cells and the basol lamina the matrix which are again always done from integrants and so we'll get back to integrants in a minute.

[00:18:53]So here is that's where I'm going to expand upon upon this the idea of when you lose ecadherin you're not just losing a cell cell connection you are changing what is happening in the cell ekadherin is involved in a cell um cell signaling process with with the conjunction of betain okay and what we're going to see and I'll kind of describe is when ekadhirin is present and connecting cells This molecule called beta betaine is in the cytoplasm. Okay, part of this kind of structure here. But what we're going to see is the loss of ekadherin will result in betain transllocating to the nucleus acting as a transcription factor and increasing everything you know that's involved in in basically cancer progression. Again, you know, it's a transcription factor many uh affecting all those different features including proliferation, migration, etc. So again, we kind of kind of expand maybe picture-wise on this as well. So just for fun, just because I have some pictures here.

[00:20:20]So again, when you have eadherin, actually I'm going to do it. I should make it a little bit bigger.

[00:20:32]Since I'm making pictures don't ask why this again just for help to illustrate this and I just think it's it's quick pull to sort of do. So again you have connections between eadhirin As long as that's the case, betaine is in the cytoplasm.

[00:21:51]However, as part as this idea of when you lose beta, I'm sorry, lose I'm just just drawing here again.

[00:22:13]So loss of eco adherent.

[00:22:27]Now we have had a better run.

[00:22:40]There's a kina.

[00:22:43]basically transllocates the nucleus and because it's a transcription factor it will end up increasing everything that you would want for uh uh for cell proliferation more secretion of MMPs etc. So again loss of ecoadherent is also then as far as that goes basically transllocation of betaine to nucleus where now it acts as a transcription factor.

[00:23:43]A similar idea then when we talked about about HIF or Hif 1 alpha in which it's in the cytoplasm for degradation but under those hypoxic conditions it transllocated to the nucleus and then acted as a transcription factor mostly for for veg everything's always a little irritating.

[00:24:11]Okay, so just a different picture of it.

[00:24:13]Okay, I don't care what pictures you like. This is a more uh scientific picture showing the idea of ekadherin.

[00:24:20]Okay. And basically it is connected though to this idea of a beta in state but the loss of it so you're losing it is basically then transllocating of the nucleus and again with some other co-actors acting as that transcription factor again part of this EMT state loss of eadherin but correspondingly you do want to get remember this idea that it's not just a connection you're not just losing cells connection you're basically changing the cell signaling again to basically promote you know basically factors involved in that we mentioned for EMT and just showing you that progression.

[00:25:12]Okay, again cool cool pictures come from this book basically putting it in and you can actually see that within a given tissue when you're stating for again ecadherin which is in green beticatina in red uh as you actually go from um from epithelial to epithelial to meenal you can actually see again where you normally had sort of this you know ecoadherent expression sort of on the outside you start to then see this transition of the loss of eucatarin in green and then you have this sort of spread of uh of betaine from where it was localized kind of outside okay near near the border of the cell localized to the cytoplasm and getting into the nucleus. So you can see within one tissue sort of this epithelial to meenal transition.

[00:26:08]Okay. So loss of be coherent when you're losing that. So the second thing and I'll put up over on the right will be this idea of again changing uh the uh the interaction with the basement membrane. And so do that I'm going to also take this Okay. And so we'll kind of put it here.

[00:26:52]Uh I keep going back and forth still to do this. I think it's optimal.

[00:27:07]kind of put on this. And so again, now we have this idea of the idea of the base membrane.

[00:27:34]Okay. So again this represents the basement membrane which consists largely of lamin protein and as we'll recall we'll put it in when we're talking about that uh ekadherin was sort of joining the cells together we had this idea of integrants being involved in basically cell matrix interaction and so we're going to degrade the laminant. So basically again we've decreased uh cell detachment okay we decrease that and then we have this idea of decrease in oops decrease in cell basement membrane uh interaction mainly by again degrading ing the major component of lamin.

[00:29:00]So the concept then of the loss of cell cell get rid of eadherin and then again we're going to be losing cell matrix by getting rid of mainly laminant proteins.

[00:29:24]So a quick picture showing this Again, just cool pictures on these things. So, kind of get that down, too. Again, showing this kind of comes from this u large book of of cancer. I call the big book of cancer by Weinberg, which I don't make people buy, but they have some cool figures, you know, showing that concept of this decrease. And so, you have again normal epic tissue surrounded in that lamin basement membrane. You could see as you're going from again epithelio to meenal you have this degradation you know and loss of u and loss of laminant loss of the basement membranes and you can actually look at a kaplan sort of meer type curve uh and basically the you look at the uh the the idea of the pro in this case looking at the probability of forming metastasis so a little bit different than kapla meer but basically the more uh the more loss of the basil membranes the more likely it is to metastasize. So just clinical data sort of supporting that idea.

[00:30:43]Okay. And again, you know, there's different, you know, this is basically really all I'm using this for is to again to illustrate kind of the figure I had up here where you've talked about loss of eadhirin and of course we we talked about what happened to betain but also now the loss of the basil lamina again the loss of laminate proteins which now allow us to get to the next stage of getting to the extracellular matrix.

[00:31:10]So I want to remind us again now about integrants and this is well I'll just keep it on this picture right now because moving it. So we we had in the we literally did this uh when we did angioenesis looking that when we look at inorgan binding so we talk about cell matrix binding cell to different ECM components they're over a class of integrants and normally being bound to it is not just adhesion it is also going to be you know activating signal pathways that we've studied to uh you know to allow it in case what survival proliferation and potentially being able to migrate along this matrix Okay. So now that we've changed the basil lamina, okay, and this is again what I'm going to want to talk about. As long as we've changed the basil lamina, now uh we're going to have different we're going to have uh different proteins uh different integrants associated with the new matrix components. And there are a bunch of other ones, okay, besides uh you know the ones I want us to know. But since alpha v beta 3, alpha beta 5 were the ones that uh I sort of had you focus on for androgenesis, we will use that as well. So it is a little variable, but these seem to be consistent. So again, you're going to have Okay. And really, I guess I really put that with this if I can which this thing is such a pain.

[00:32:52]two.

[00:32:55]Okay. And so now we also then have this idea of you know a change of integrance and specifically looking at again for us to know an upregulation of alpha v beta 3 upregulation of alpha v beta 5 on that too. So again, now we're switching again to these guys. Alpha V beta 3, alpha V beta 5.

[00:33:38]Okay, so again you have this idea starting out. You basically you got loss of of uh of eadherent for those cell connections. You have a loss of basement membrane by getting rid of the lamin. And you've had this upregulation of new integrants alpha beta 3 alpha 5.

[00:34:09]That's going to allow us then to continue the journey through the matrix.

[00:34:14]And let's we'll skip the other besides these MMPs we're going to talk about.

[00:34:17]We're going to skip for our purposes this other uh important uh proteiase. Uh but again we don't need to know everything for this.

[00:34:27]Okay. So and again I guess I put this put this as number three because I'm really just going through sort of the steps here. So you have change of integrance and again you have that's upregulation of these MNPs and other proteases. is okay upregulation of MPs. And now we're going to start degrading the matrix.

[00:35:13]Okay, which is kind of this part as I have here. And again, it's going to be localized. When you look at this picture, we're not going to just produce MMPPS that are just everywhere along here. We have a path just like we sort of had a path for angioenic vessels going up, we have a path for these cancer cells going down.

[00:35:36]Okay. And again, this is just this idea of when you look about uh you know the concept of this again, you're going to have this migration. And when you think of migration now and again I'll show some pictures of it. Again these cells are not just floating you know floating through here they have to basically crawl along cells I mean crawl along matrix and it's going to be again this dynamic process of this. If you look at the matrix that it's crawling along to, again, you sort of are going to have cells that are again going to be meenal like and they're going to have to again sort of stretch.

[00:36:36]They're going to sort of bind by.

[00:36:40]But in order to move, they're going to have to then be attached back there. So then they could slide forward and do it again where they're going to extend the side of skeleton attached by integr so they can go forward. So it's a dynamic process. So upper relation of MPs and then it's this dynamic motility.

[00:37:18]Okay, shown by this again kind of basically just saying that right as I just said this and again here's some sort of pictures some time lapse of some cells you can actually watch online of actually moving but literally you sort of see the spread of moving forward you know this is lamelopolia sort of attaching in these kind of adhesions here and then you can see they put stars sort of detaching and back so now the cell that was here already is further down and that's sort of what happening across the board here.

[00:37:50]Here's just some other pictures of this uh some just cooler pictures of showing of showing again just for fun the the the extension of the metapolium. Okay, from two different angles here.

[00:38:06]Okay, so this is important as far as how they are migrating these epithelial cells. And this is really cool. The idea is they're not migrating as single cells. Okay, here I sort of had them as a single cell going to a single cell.

[00:38:21]They're not doing that. They're going as cohorts for the most part for these carcinoma type cells. And the idea of that is going together as a cohort. The cells at the front are going to be involved in having the most amount of these integrants to involve sort of that adherence and also MMPs. So they're sort of presenting more of make making a hole this direction versus just everywhere else. So they're in charge sort of of the lead. They're going to be making some MMPs to dig their way through.

[00:38:50]They're are going to be the main ones sort of, you know, with those strong in binding to bind. And then in the back you're going to have cells which under different signals. So the signaling through the cells will cause sort of a weakening of the uh of the inrine bind bonds. And so that will allow them to this detach and move forward.

[00:39:13]Okay. Not only again this is this idea of how they migrate. Not only are they doing it as far as being leaders, they also get basically stromo cells. Okay. And so the idea of in this case they've recruited their their carcinoma associated fiberblast to lead the way to basically you know produce the MMPs etc and basically make the path for them and so here's you know the the way of invasion.

[00:39:42]So you have this idea of a cohort of cancer cells but here also you see using uh sort of the stroal cells that that the cancer is controlling to help him along the way too.

[00:39:53]I think that is totally cool.

[00:39:57]Okay, so those are the early steps and again we'll have this all on one on one page. Let's kind of get to the last few steps as well. So again, can't put I have this on one page later, but let's sort of erase this and get the last few steps that I want to talk about as far as the metastatic cascade. And then we'll have one slide that I sort of made is probably the key one for us to get out of this.

[00:40:34]And of course now I've got to like spend time erasing this.

[00:40:40]You can move forward in your little tape.

[00:41:23]See, do I have a good red one here?

[00:41:49]Okay. So, we'll go on to four or as I'll describe four again. This is introversation.

[00:42:09]Okay. Now this is going from basically extracellular matrix into the blood.

[00:42:24]That sort of introversation entering the blood. And again the cartoon version and this will give of course will give uh some details to us. The cartoon version of this is okay how are these cells okay they're a little more clumped they probably will be how are they going to get out of this cell matrix you know into or you know the extracellular matrix uh into the blood vessels. Well in a lot of cases it's it's basically and we'll show you we'll show you that those instances the details it's going to be these endothelial cells retracting. So these cells can slide in between and then in general again they'll come back together and again the angioenic vessels already are considered leaky okay because they weren't formed properly. So that kind of helps them get between cell cells but there's also a mechanism okay if you think the mechanism is nothing that cancer is going to invent. Again when you think about uh immune cells that are circulating they have to get in tissues they have some of the same features that are involved in causing these cells to retract.

[00:43:26]But when we look at this, I'm going to be breaking these up in a couple slides on this too. We have to sort of do two things. We have to when you look at the connections between u endothelial cells, there is another type of caderin called you know vascular endothelial or vheadherin. So it's going to be sort of two steps here involved in in sort of this process here. You're going to have to cause breakup of these. Okay. So, disruption or getting rid of those ve and then you have to cause the cytokeleletal of these endothelial cells to to u you know to move to change so that then they retract from one another the cancer can go through. So again when you're looking at conversation for us you are looking sort of at and again I'll do it the way I have it here.

[00:44:16]You have basically Duption disruption of the VE cadherin which again is going to be what's connecting these cells are these are our famous end the ilio cells and then you're going to have to again cause retraction of these cells.

[00:45:20]So the cancer cells here's our cancer cell can migrate through now we will be in the blood. So as far as disruption of vaderin again and we sort of have this here you're going to have basically these guys secrete the cancer cells secrete these MMPs.

[00:46:00]But these MMPs actually bind to again what are what I can kind of see there on this too. Basically these you know PAR receptors which again are basically being activated by these proteins.

[00:46:16]PAR receptors PAR receptors on the anilio cells.

[00:46:28]Okay. And again, that's the main thing that's gonna that's going to help to break up these bonds. Okay. So that's disruption of the Vadhirin. And then the second part.

[00:46:41]Oh, endthelial cell retraction which again now we will have this idea of who's going to help us out. It's going to be a tumor associated macrofase. So we're going to get the help of the tumor associated macrofasages which are here.

[00:47:10]And again, they're going to release TNF alpha that again and you could sort of see it releasing TNF alpha. You're kind of doing it's going to bind to receptor which we don't have to name.

[00:47:45]to trigger retraction.

[00:47:59]So again if you look at your tumor surge macridges now they are secretreting then TNF alpha but bind again that's really involved in retraction. So again two steps I sort of want you to know you again they are still simplified but the idea even to go back to the slide that you're going to have to have uh this idea of this is all introversation first you have to disrupt these basically vadherin that's basically MMP's binding to the par receptor okay that's going to help break these down and Then you have the tumor associated macrofasages shown here in my little diagram here releasing TNF alpha. It binds to receptors on the endothelial and cause retraction and cells can get in between the cells. Okay, we're not going to discuss the way that that cancer cells can also get in you know via through the cell via endoccytosis exocytosis involves calcium. We don't really care.

[00:49:20]So this is kind of the step of interposation that I sort of care about.

[00:49:23]break up of the uh Vadhirin and then uh endothelial cell retraction with a couple details here but you can also see again cancer cells are not doing this alone they're controlling or they're controlling macrofasages to actually help produce factors they need as well so let's just throw this picture here and get this concept cept call it again tumor micro environment of metastasis how much do we have to say tumor micro environment to sort of kind of get it together and so and it's sort of clinically significant so they can actually look again macrofasages the tumor cells and ailio cells forming this triad called tmem and again it has you know this is sort of it the more of these you have and you can quantify it the it's basically you know negatively correlated with survival the worse worse prognosis.

[00:50:27]So again, you have this concept of I have another pen, it might not work, but you have tumor micro environment of metastasis.

[00:51:04]Again, it's the tumor associated macrofasages.

[00:51:08]It's the cancer cell and it's the endothelial cell. this this triad of these three bad significance or bad prognosis and you can see them here.

[00:51:21]So ination not that easy but we've made it there.

[00:51:37]Okay. So how about survival?

[00:51:41]Okay. So again, and we'll see this in the next lecture too. Cancer cells are not supposed to be are not supposed to be in the the bloodstream which make them which make them susceptible to immune cell attack and just the fact of the physical rigor of going through blood vessels and capillaries. Again, they are not designed to do so. So, they've got to come up with a way to survive.

[00:52:29]And so, they have a clever way of doing it.

[00:52:43]And that is Let's make a nice little picture bigger.

[00:53:05]for this. So we have survival.

[00:53:11]Okay.

[00:53:21]Survival and blood puts five again what the cancer cells are going to do. So here is they're going to surround themselves with platelets.

[00:54:02]which again accomplishes the two things I already mentioned. It basically protects from immune cells and sort of stabilizes the journey.

[00:54:24]Kind of takes up the shock of banging around these these blood vessels.

[00:54:37]Again, complicated picture. It comes from an article I like got to be an easier way to do this too by the way on the again just to show the picture sort of bigger on this so that we'll see it again here's our circulating tumor cells okay they get clustered again by by the u by platelets and again sort of this idea of protecting from the these innate immune cells and protecting it from stress us until we get to later on we're about to get to this idea of the second phase which will be them adhering and then getting into the new place. So we're just concentrating on this slide uh for this part how they protect themselves, how they're surrounding, how they survive, right? So okay so here's the idea of and this is where it's not completely known too okay they're circulating into the system but why do metastasis go to certain places why do they arrest in particular capillary beds uh and you know proceed we're going to see the next lecture sort of how that is uh sort of studied here Okay. Are there specific markers in capillaries of u of target organs that cancers normally go to? Are they simply being trapped because as we'll see next time the the cells themselves cancer cells are bigger than the capillaries.

[00:56:13]Okay. So we'll kind of visit that but basically simple idea of one way or the other they have to be you know arrest or be trapped in the capillary bed of some sort of tissue.

[00:56:36]Again we'll talk about mechanism in the next lecture.

[00:56:51]So, the last real one and kind of we'll look at this and may make it in a couple bullet points.

[00:57:00]If I can just I wish I could do this easier because it's frustrating on this thing too. Again, complicated slide. We're not going to have to know all the details. I'm just giving certain details of it. This just came from a paper again, but this is going to be cool. We're going just put a couple words on this too. The fact is, you know, they're they're going to basically adhere. A lot of times they're going to adhere by the platelets themselves.

[00:57:25]Okay? And as we're going to see, I'll put in words by after adherence, the cells are going to communicate to the endthelial cells. And who are they going to get? They're going to get some help from some of these innate immune cells that we've talked about before.

[00:57:43]Okay. So, let's actually just have me look through this instead of even writing it down.

[00:57:50]It's pretty pretty well attached here.

[00:57:51]Trying to use again just pieces of this article that came out. And so, basically, again, platelets are going to attach a lot of times. You know, the cell is going to attach via these platements uh attachment. You know, platelets are designed to send out molecules to recruit other other types of cells. A lot of times platelets for forming clots here, what are they going to do? They're going to get neutrfils.

[00:58:16]And this is really cool right here.

[00:58:18]They're going to have neutrfils uh basically attracted to the site. And this is amazing. They're going to release uh these soluble DNA. And what they're going to form are basically called nets to help to tra to help trap some cancer cells here as well.

[00:58:35]So again, platelet attachment of cancer cells, platelet recruitment of neutrfils and neutrfils definitely secretreting all kinds of factors including DNA to form these nets that actually help attract u help attract and and bind to uh cancer cells, which is crazy.

[00:58:58]And then again simplifying what a lot of these details are. You're also going to recruit monocytes. Monocytes actually become macrofasages. And monocytes are going to release different things that I don't feel like memorizing and veg upf.

[00:59:12]But what you're going to end up doing now is instead of creating let's say you know tumor associated macrofasages they're sort of going to be a subclass be called metastasis associated macrofasages.

[00:59:24]Okay, these are basically then going to secrete things which are more specific for this extraation which we won't go into the details of it but but different things on this. So again you have this process here of these three steps of platelets uh cancer cells are attaching to endothelial cells via platelets.

[00:59:45]Platelets are sending off factors that will recruit some of these other cells some of these natural immune cells. this case neutrfils we're talking about neutrfils release DNA to form these nets macrofasages are going or I'm sorry monocytes uh which are the precursors to macrofasages they're going to involve in the secretion of substances that will convert them not into what we're calling tumor associated macrofasages but more specifically metastasis metastasis associated macrofasages mam which simply will secrete different substances to help this this secondary process here of metastasis as far as eight goes which is mainly the next lecture and then we'll put this summary slide up is going to be again you know in order for the secondary growth of this to occur you need to have a suitable micro environment okay and then once that happens you could go through the entire process again but you have to I put in bold because that is the main focus of it because what we'll describe next time and this will make more sense. We'll actually start here. If you just look at the the combinations as far as the suitable micro environment first of all it might already exist. Okay, the seed the cancer might match the soil. Okay, the different growth factors and things that are already there. So it might already be there. Okay, other other times cancers won't die but they won't grow very well. they'll wait for factors to change. Okay? Maybe then you'll have more, you know, more exposure to carcinogens, different things will happen, hormones, etc. that will then have it where the environment will be produced. But cancer cells don't like to wait. They like to take charge. So, we'll give an example next time where they actually once they get there, they'll create the proper micro environment. They'll remodel once they're there to get what they need.

[01:01:50]are and this is sort of this idea of which is you know which is that is the idea that sometimes they modify the micro environment before they actually arrive. Okay, they create what's called a pre-tastatic niche. And so much of the next lecture really will be talking about how cancer does all of this because this is the key again even though it may be able to at that point to already get to the point of uh of these last steps of you know where it's actually again arrested in the in the organ here made all the way here the idea is and even extra visated this is going to be the key does the micro environment exist or can it be created.

[01:02:35]If you could stop here, poison the soil so it doesn't move on. It doesn't matter if the cancer cells are necessary there, you'll die of something else.

[01:02:46]So again, this is something I made which is cool. At least I think that we'll be able to see in class as well.

[01:02:56]I wish I could make it a little bit bigger but we can kind of see it kind of again putting this picture but putting all some of the major details in one place since there was all sort of complication here again is looking at cancer cells uh under normal state before they metastasize here they're they're connected by ekadherin they're connected by particular introins to the basement membrane which is lamin okay so decrease increase in cell cell interaction. We've what is this thing?

[01:03:33]We've again basically lost eadhirin.

[01:03:37]We've gained actually enh here because that's different. I made the cells started to be meenal like okay you degraded the matrix. You decreased the the uh cell matrix by degrading the lamin. That's shown by the dot dot dot.

[01:03:52]Okay. You're migrating through again the is you're producing MMPs and now you have these new inens I made them red alpha v beta 3 and beta 5 they're going to bind upon different uh new extracellular matrix com you know components that are along the way now as they're producing them because MMPs break down the matrix but they leave fragments and these are what the cancer cells are going to basically migrate along so you have that okay intravisation getting through the blood vessels. I tried to again this this is supposed to represent Vadhirin, but I kind of wrote this here, right? You're going to degrade Vadhirin and you're going to trigger basically uh retraction of those cells. Boom. And those cells retract and retract. Five, intro. I have them, this is cool. I like the way I do this. Just surround themselves with platelets. Again, that protects them from lymphosy attack.

[01:04:51]that's that's sort of in the circulatory system and also the fact that these are going to have to migrate through some very small capillaries and things and be bouncing against the walls so it protects them.

[01:05:03]Okay. And then seven and I kind of put since that was a complicated sort of slide here too kind of put most of the steps right here. Extravisation is leaving the bloodstream and getting uh you know into the into the target tissue. Again the idea of platelets I kind of showed here platelets are attaching the cancer cells with them they're recruiting neutrfils again kind of wrote this to form these nets if I really made my net really good and also platelets recruit monocytes which will end up becoming uh you know macrofase metastasis associated macrofasages they kind of get it here and then we have this okay if the conditions are right okay to make it all the way through here you basically You want to reestablish this epithelial basically phenotype that you started with here again epithelial meenal qualities epithelial again if this environment is right. So that's sort of the metastatic cascade is in at least a few steps but you really see the dynamic process that has to occur and we're manipulating all kinds of cells normal cells along the way changing the matrix as well. Okay. So you can just sort of see some things as far as you know controlling beta and I talked about you know inhibiting these MMP production having particular inocrine inhibitors which some people will talk about poisoning these environments so it is not conducive to migration either way and again this idea of well this is sort of an interesting idea here if platelets are surrounding it okay and platelets form clot maybe clot breakers or some form of it may be able to be used to decrease this the although again it might decrease clotting as well but it's sort of an idea each step along the way you want to think about what combination of therapeutics you would apply to counter all of these moves that the cancer is making next time we'll specifically look at poisoning this environment making it where it's not conducive for cancer growth then it doesn't really matter if a couple of those cells made it out here and didn't do anything uh the idea is you You'll be able to live, you know, basically fine if you can just keep that at bay.