WEBVTT - How Gene Therapy Works 0:00:04.280 --> 0:00:07.240 Get in test with technology with tex Stuff from how 0:00:07.320 --> 0:00:14.760 stuff works dot com. Hey there, we one, and welcome 0:00:14.800 --> 0:00:17.800 to tech Stuff. I'm Jonathan Strickland and I'm luring voc 0:00:17.840 --> 0:00:21.800 Obama and today we've got our topic courtesy of a 0:00:21.920 --> 0:00:26.120 listener request. Yes, this is listener Mike via Twitter. I 0:00:26.160 --> 0:00:29.000 didn't write down his screen name, so one of the 0:00:29.040 --> 0:00:31.840 Mike's on Twitter, thank you sir yea and said, have 0:00:31.880 --> 0:00:34.240 you done a podcast on gene therapy? Is that something 0:00:34.280 --> 0:00:37.519 that at Blow the Mind would do instead? That's a 0:00:37.880 --> 0:00:41.280 stuff to blow your mind. That's the Julie Robert Julie 0:00:41.320 --> 0:00:45.360 doing their their science podcast, and we really we've talked 0:00:45.400 --> 0:00:48.800 about gene therapy a little bit on Tech Stuff in 0:00:48.840 --> 0:00:51.000 the past, we've never done a full episode on it. 0:00:51.000 --> 0:00:53.720 We've also talked about it on a sister podcast, the 0:00:53.760 --> 0:00:56.640 Forward Thinking Podcast all right, along with Joe McCormick. We 0:00:56.640 --> 0:01:00.160 we did a really great introduction of what jeans are 0:01:00.200 --> 0:01:02.600 and some of the really interesting cutting edge stuff going 0:01:02.600 --> 0:01:04.920 on in gene therapy. So if you would like to 0:01:04.920 --> 0:01:07.119 listen to that, we we highly recommend it. We think 0:01:07.120 --> 0:01:09.040 that we did a pretty excellent job. Yeah. Yeah, the 0:01:09.080 --> 0:01:11.399 fact we had Joe there too. You know, put facts 0:01:11.400 --> 0:01:13.400 in between the jokes that I made and the shaking 0:01:13.440 --> 0:01:16.160 of Lauren's head. It really was different from a tech 0:01:16.200 --> 0:01:19.200 stuff episode. No, of course, I'm proud of all the 0:01:19.240 --> 0:01:21.320 work we do, and then one was a great show. 0:01:21.600 --> 0:01:24.080 But we're going to give an overview of what genes 0:01:24.080 --> 0:01:29.600 are and some of the actual technical ways that scientists 0:01:29.640 --> 0:01:32.680 and doctors are working with gene therapy right now, because 0:01:32.680 --> 0:01:35.680 in the forward Thinking episode, it was more about the applications, 0:01:35.760 --> 0:01:37.520 right it was more about why you would use it 0:01:37.760 --> 0:01:40.600 and not how like how has it actually done? And 0:01:40.640 --> 0:01:44.160 it's kind of fascinating. Yeah. Absolutely, and and this is, 0:01:44.200 --> 0:01:45.800 by the way, one of those things that we kind 0:01:45.800 --> 0:01:47.480 of struggle with all the time of of you know, 0:01:47.520 --> 0:01:49.280 we'll we'll come up with a concept for a show 0:01:49.320 --> 0:01:51.720 and then go, how should we check with car stuff? 0:01:51.760 --> 0:01:53.760 Should we check with mind? Should we you know? Who 0:01:53.760 --> 0:01:55.520 should we ask? And make sure that we're not treading 0:01:55.520 --> 0:01:58.240 on their toes and re copying ourselves because we already 0:01:58.240 --> 0:02:01.880 did a forward thinking episode and really the answer is always, um, 0:02:01.920 --> 0:02:05.080 if it's interesting and there's more avenues within a topic 0:02:05.120 --> 0:02:07.360 to talk about, then then we'll go ahead and cover it. 0:02:07.480 --> 0:02:11.600 So let's start with the very basics. And but to 0:02:11.680 --> 0:02:13.639 start with the very basics, we've got to look at 0:02:13.720 --> 0:02:18.680 cellular biology. So anyone who was currently in middle school 0:02:19.040 --> 0:02:20.920 is going to be able to go on about this 0:02:20.960 --> 0:02:23.799 at length. I actually had to go back and look 0:02:23.880 --> 0:02:26.560 up all this information because middle school was a long 0:02:26.600 --> 0:02:29.680 time ago for me. And while I was originally interested 0:02:29.720 --> 0:02:34.120 in going into biology, yeah I was, Biology was number one, 0:02:34.240 --> 0:02:37.240 and then it's always terrible most mostly that was not 0:02:37.280 --> 0:02:40.120 my branch of science anyway, Biology and physics I love them. 0:02:40.120 --> 0:02:41.880 And then I got into college and I switched to 0:02:41.919 --> 0:02:44.760 English literature with a focus on Shakespeare, and now I 0:02:44.760 --> 0:02:49.040 talk about technology. So I mean it's a clear pathway. Yeah, 0:02:49.200 --> 0:02:54.840 logical progression so sells, you know, sells the basic little 0:02:54.960 --> 0:02:58.480 unit of life for for us, we human beings, and 0:02:58.560 --> 0:03:00.760 most of the stuff we tend to interact with that 0:03:00.919 --> 0:03:05.160 isn't super teeny tiny um cells have what is called 0:03:05.200 --> 0:03:08.400 a nucleus. It's kind of the brain center of a cell, 0:03:08.480 --> 0:03:11.120 the control center if you prefer, and they have a 0:03:11.160 --> 0:03:14.320 cell membrane, and both of these things are incredibly important 0:03:14.360 --> 0:03:18.360 with gene therapy. So within the nucleus you've got chromosomes, 0:03:18.360 --> 0:03:22.399 and chromosomes are made up of two things, DNA and protein. 0:03:22.880 --> 0:03:25.919 And the DNA is the that's sort of the very 0:03:25.960 --> 0:03:31.000 basics of what makes you you. And along those chromosomes 0:03:31.000 --> 0:03:34.520 are little sections of DNA that we call genes now. 0:03:34.560 --> 0:03:38.920 The genes their job is to code for the production 0:03:39.120 --> 0:03:42.760 of particular proteins, and that's what we call a gene 0:03:43.200 --> 0:03:47.000 expressing itself. When a gene is expressed, it's coding for 0:03:47.120 --> 0:03:51.480 a protein. And that's also important because with gene therapy, 0:03:51.640 --> 0:03:55.200 one of the goals is to introduce new genes into 0:03:56.120 --> 0:04:00.040 a cell so that a particular thing is express a 0:04:00.280 --> 0:04:04.520 particular protein is created, and it doesn't always work right. 0:04:04.640 --> 0:04:08.280 So and the reason for that is because gene therapy 0:04:08.320 --> 0:04:12.160 this is still a very young science and uh and 0:04:12.520 --> 0:04:17.120 sometimes the techniques we use have high success rates, sometimes 0:04:17.120 --> 0:04:19.800 they have low success rates, and so it may be 0:04:19.920 --> 0:04:23.600 that you're doing everything right but or as right as 0:04:23.640 --> 0:04:26.480 we know how to do right now, but it's still 0:04:26.520 --> 0:04:30.800 not you know, success rate right since it's so experimental. 0:04:30.920 --> 0:04:33.560 Right now, we're currently on using gene therapy to um 0:04:33.640 --> 0:04:38.760 to treat otherwise incurable diseases and particularly childhood diseases right 0:04:38.920 --> 0:04:42.479 and only in clinical trials for that matter, right. You 0:04:42.480 --> 0:04:44.520 you have to be part of a clinical trial, and 0:04:44.720 --> 0:04:46.839 the only gene therapy that is going on is being 0:04:46.839 --> 0:04:49.920 conducted on somatic cells. Those are those are body cells 0:04:49.960 --> 0:04:53.760 as opposed to reproductive cells, right, which means that any 0:04:53.880 --> 0:04:56.800 changes that are going on, and and the kinds of 0:04:56.880 --> 0:04:59.800 changes that we're making are either inserting normal genes for 0:05:00.040 --> 0:05:03.840 herring abnormal ones or altering the kind of on off 0:05:03.920 --> 0:05:07.200 switches of those genes. Um they're only going to affect 0:05:07.200 --> 0:05:10.520 the patient, and any hypothetical offspring of that patient would 0:05:10.520 --> 0:05:14.200 not would not would not inherit those those genetic changes, 0:05:14.240 --> 0:05:18.240 because again they're somatic, not the reproductive cells. So in 0:05:18.320 --> 0:05:21.320 another issue would gene therapy. Similar to that is that 0:05:21.720 --> 0:05:26.040 often the benefits that you experience with gene therapy are 0:05:26.080 --> 0:05:29.160 temporary in nature, and that you have to undergo multiple 0:05:29.279 --> 0:05:34.200 gene therapy sessions to have any lasting effect because they 0:05:34.279 --> 0:05:39.839 don't necessarily uh change the genome. It doesn't necessarily change 0:05:39.880 --> 0:05:43.800 everything like the actual uh way the cells are forming 0:05:43.839 --> 0:05:46.760 from that point forward in your body. Right, It might 0:05:46.800 --> 0:05:51.080 just affect the current expression of that current cell, that 0:05:51.120 --> 0:05:53.599 current gene within that current cell, and if that cell dies, 0:05:53.960 --> 0:05:56.880 then you know that's the the benefit you got from 0:05:56.880 --> 0:06:00.160 that one cell is over. Yeah. I'll also, genes are 0:06:00.160 --> 0:06:04.080 really complicated. They most of them make more than one protein. 0:06:04.120 --> 0:06:06.960 That average is three, which is something that we've learned 0:06:06.960 --> 0:06:09.599 from the Human Genome Project, which I'll be talking about 0:06:09.600 --> 0:06:12.880 a little bit later on. It just means that, you know, 0:06:13.160 --> 0:06:16.880 genes don't have they're not self contained bits of DNA, 0:06:17.040 --> 0:06:20.359 and they don't have strictly defined roles. Yeah, so you 0:06:20.440 --> 0:06:23.400 can't just say just get rid of that one, right, yeah, 0:06:23.400 --> 0:06:27.240 because it could be that a disease is actually the problem. 0:06:27.320 --> 0:06:29.559 You know, it might be the product of a few 0:06:30.000 --> 0:06:31.920 different genes, and then you have to figure out, well, 0:06:31.920 --> 0:06:35.080 which one of these genes is not behaving the right way, 0:06:35.440 --> 0:06:37.200 you know, which one is the mutated gene, which one 0:06:37.200 --> 0:06:39.839 do we need to replace? And then by replacing that 0:06:39.920 --> 0:06:42.520 gene are we causing any other issues? Is there some 0:06:42.600 --> 0:06:45.719 other problem that could happen as a result of going 0:06:45.760 --> 0:06:49.240 in and altering a person's genes. And this is, like 0:06:49.279 --> 0:06:51.719 I said, a very young science. So we're still learning, 0:06:51.760 --> 0:06:54.000 and that's one of the reasons why clinical trials are 0:06:54.000 --> 0:06:56.560 the only place, at least in the United States where 0:06:56.600 --> 0:07:00.120 you can get gene therapy and you know, because honestly 0:07:00.240 --> 0:07:03.600 it's not ready for a full rollout yet, certainly not now. 0:07:04.080 --> 0:07:07.719 So here we've got a cell, we've got that cell membrane. 0:07:07.960 --> 0:07:11.520 By the way, DNA does not pass through cell membranes. 0:07:11.560 --> 0:07:15.120 Cell membranes are semipermeable, right, which means that they allow 0:07:15.360 --> 0:07:18.720 some stuff through and they allow they keep other stuff 0:07:18.760 --> 0:07:23.400 either inside or outside. Now, DNA generally speaking, is something 0:07:23.400 --> 0:07:26.440 that cells want to keep inside. They don't like letting 0:07:26.480 --> 0:07:29.320 the DNA go flying out all over the place. It's 0:07:29.320 --> 0:07:32.000 a better idea, yes, So that means that if we 0:07:32.040 --> 0:07:34.720 want to introduce DNA into a cell, we have to 0:07:34.800 --> 0:07:37.960 kind of find a way of getting the DNA through 0:07:38.000 --> 0:07:42.040 the membrane without damaging the cell because clearly or the DNA, 0:07:42.280 --> 0:07:44.600 So if you kill the cell in the process, that 0:07:44.760 --> 0:07:46.720 that's not what we call youthful. We call that. We 0:07:46.800 --> 0:07:50.560 call that a fail that's that's generally speaking, that's a failure. 0:07:50.800 --> 0:07:53.280 You don't you don't want the cell to die. So 0:07:53.760 --> 0:07:57.480 finding a way to get DNA material into a cell, 0:07:57.920 --> 0:08:01.560 So getting a good gene through that cell wall without 0:08:01.640 --> 0:08:04.880 killing the cell has been really the main focus of 0:08:05.560 --> 0:08:09.880 genetic therapy over the last couple of decades. So there 0:08:09.880 --> 0:08:11.600 are a lot of different ways of doing it, and 0:08:11.640 --> 0:08:14.680 we could either go about this chronologically or we could 0:08:15.280 --> 0:08:20.120 try thematically or maybe alphabetically. But we kind of decided 0:08:20.200 --> 0:08:21.920 like the first one we wanted to look at is 0:08:21.960 --> 0:08:26.320 in a way, sort of the simplest, at least in concept, 0:08:26.440 --> 0:08:29.760 if not in practice. And I guess the we're we're 0:08:29.760 --> 0:08:34.559 going to cover two basic categories UM in this podcast 0:08:34.559 --> 0:08:37.120 today and a couple of sort of outliers and and right, 0:08:37.160 --> 0:08:38.960 there's a bunch more. We'll talk about that a little 0:08:38.960 --> 0:08:40.360 bit at the tail end of the episode, and I 0:08:40.400 --> 0:08:44.520 guess that we're sort of going chronologically a little bit. Yeah, 0:08:44.559 --> 0:08:47.080 it's a it's a little bit of a jump around. 0:08:47.120 --> 0:08:49.600 But but the first one we wanted to talk about 0:08:49.840 --> 0:08:54.000 is micro injection, which is more or less what it 0:08:54.040 --> 0:08:57.640 sounds like. Yeah, you're talking about a very very tiny 0:08:58.720 --> 0:09:02.760 needle going into a cell and injecting DNA material directly 0:09:02.800 --> 0:09:06.840 into either the cytoplasm, which is the the stuff inside 0:09:06.880 --> 0:09:10.520 a cell membrane just in general. That's that's the uh, 0:09:10.640 --> 0:09:13.200 both the fluid and all the little organelles and everything 0:09:13.200 --> 0:09:15.840 that are in a cell, or you're injecting it directly 0:09:15.880 --> 0:09:18.160 into the nucleus, which is where the d n A 0:09:18.280 --> 0:09:21.080 material is kept. So in other words, you are using 0:09:21.280 --> 0:09:24.760 a needle and you're just inserting the DNA directly there. Now, 0:09:25.080 --> 0:09:28.360 when you're talking about cell cells are I don't know 0:09:28.360 --> 0:09:31.400 if you know this, they're tiny. They're they're really little. Yeah, 0:09:31.480 --> 0:09:34.840 you might think that, uh, the needle that you're getting 0:09:34.920 --> 0:09:38.200 for techna shot is huge. Well it is pretty big, 0:09:38.240 --> 0:09:42.440 but to a cell, that's gargantuan and I'm nearly positive 0:09:42.480 --> 0:09:45.840 that many cells could fit inside. Yeah, so you need 0:09:45.880 --> 0:09:49.000 to find a very very fine needle and you have 0:09:49.080 --> 0:09:51.280 to be able to make. Really, you're not You're not 0:09:51.320 --> 0:09:54.560 gonna You're not gonna just stumble over one of those. Okay, well, 0:09:54.600 --> 0:09:57.040 I mean, you know, if you happen to stumble in 0:09:57.080 --> 0:10:01.600 a lab, you might, but I accept that you make 0:10:02.000 --> 0:10:05.000 an incredibly fine needle, and you actually you kind of 0:10:05.040 --> 0:10:07.160 need to. One of them is just less fine than 0:10:07.200 --> 0:10:08.800 the other one. The element is more of a pipette, 0:10:09.200 --> 0:10:11.400 and the reason for that is you have to immobilize 0:10:11.480 --> 0:10:14.439 the cell that you want to inject. You need to 0:10:14.480 --> 0:10:16.680 kind of hold it down because otherwise cells are sort 0:10:16.679 --> 0:10:19.480 of squid squiggly and yeah, they are. They do tend 0:10:19.480 --> 0:10:23.880 to squiggle. Okay, So so this this technique was really pioneered. 0:10:24.240 --> 0:10:28.360 The needle micro injection in general was pioneered by one 0:10:28.480 --> 0:10:31.839 doctor Marshall Barber in the early nineteen hundreds. UM. He 0:10:31.920 --> 0:10:35.319 was developing it to study bacteria and confirm the germ 0:10:35.440 --> 0:10:37.800 theory that was being developed around that time by a 0:10:38.679 --> 0:10:45.599 Cotchin pastor UM who would who would basically, um outline 0:10:45.640 --> 0:10:48.760 the entire germs make a sick kind of contract, which 0:10:48.760 --> 0:10:51.520 was revolutionary at the time. Right, it's still I'm still 0:10:51.559 --> 0:10:54.040 pretty glad that we have it, and I think I 0:10:54.080 --> 0:10:57.800 think it's great. So early on, like through the nineteen sixties, 0:10:57.880 --> 0:11:00.679 early on, I mean, this was a very slowly developing 0:11:01.000 --> 0:11:04.839 field of study. Uh. Microinjection was used to implant whole 0:11:04.960 --> 0:11:08.679 organisms and embryos into larger bodies. Then in the nineteen 0:11:08.800 --> 0:11:13.520 seventies we started implanting cellular organelles and molecules UM and 0:11:13.760 --> 0:11:17.160 other relatively large bits of stuff into cells. And then 0:11:17.160 --> 0:11:19.840 as we got into genetic study in the mid eighties, 0:11:19.880 --> 0:11:23.080 we began injecting stuff like proteins and DNA and RNA 0:11:23.480 --> 0:11:27.240 into cell nuclei. Right, So it obviously took quite a while. 0:11:27.400 --> 0:11:32.280 And the general process, again at least conceptually, is simple. 0:11:32.400 --> 0:11:34.800 You hold the cell still and then you inject the 0:11:34.800 --> 0:11:37.680 stuff into it. Yep um, it's it's all. It's all 0:11:37.720 --> 0:11:41.160 the same process, just just as a miniaturization of these 0:11:41.200 --> 0:11:46.480 technologies began to get gooder. Yeah, right, because because otherwise 0:11:46.480 --> 0:11:49.320 holding a cell still that's actually pretty tricky. They use 0:11:49.400 --> 0:11:51.840 these pipettes that use a little bit of suction that 0:11:52.000 --> 0:11:55.600 immobilize the cell, and then you have to have a 0:11:55.679 --> 0:11:58.960 micro manipulator that's something that allows you to make very 0:11:59.120 --> 0:12:02.200 very precise movements because you obviously you could not use 0:12:03.440 --> 0:12:06.960 a handheld hypodermic needle. You wouldn't have the precision to 0:12:07.000 --> 0:12:10.920 be able to to target specifically the nucleus with any 0:12:11.080 --> 0:12:13.240 level of confidence. I was, I was pretty good at 0:12:13.280 --> 0:12:15.280 duck Hunt, and I've got to say that I would 0:12:15.280 --> 0:12:18.000 not that I don't have that kind of precision. The 0:12:18.040 --> 0:12:22.240 hand eye coordination is pretty pretty uh remarkable. So yeah, 0:12:22.240 --> 0:12:24.440 they usually you have some sort of device that is 0:12:24.520 --> 0:12:27.680 designed to to study the needle and direct it properly. 0:12:28.480 --> 0:12:33.959 So has limitations. There are some big ones actually. Also 0:12:34.000 --> 0:12:37.320 in order to really use this technology, we we would 0:12:37.320 --> 0:12:39.559 have to start getting into them what we learned through 0:12:39.559 --> 0:12:43.280 the Human Genome Project, which um, which didn't really get 0:12:43.320 --> 0:12:47.160 started until the nineteen nineties. I mean, it came about 0:12:47.200 --> 0:12:50.880 after the Atomic Bomb Project in World War Two, actually 0:12:50.920 --> 0:12:55.280 because Congress charged the Department of Energies predecessor agencies with 0:12:55.400 --> 0:12:59.960 UM studying and analyzing genome structure, the replication, damage and repair, 0:13:00.000 --> 0:13:04.920 air and and consequences of genetic mutations. UM. So you know, 0:13:05.080 --> 0:13:08.760 especially those that were caused by radiation and the chemical 0:13:08.880 --> 0:13:14.800 byproducts of warfare energy production. Right. So, UM, the Genome 0:13:14.800 --> 0:13:17.920 Project completed its initial research a couple years ahead of 0:13:17.920 --> 0:13:23.240 schedule in two thousand three. And UM, that's basically where 0:13:23.280 --> 0:13:26.120 where we got most of our information. You know. That 0:13:26.120 --> 0:13:29.520 that that that is the spawn, that the dawn of 0:13:30.480 --> 0:13:33.520 gene knowledge, right, you know. Without that, obviously, we couldn't 0:13:33.559 --> 0:13:38.600 make any um, any knowledgeable decision about how to go 0:13:38.679 --> 0:13:40.800 about this at all. Right, we wouldn't know what was wrong, 0:13:40.920 --> 0:13:43.280 much less how to correct it, all right. And so 0:13:43.320 --> 0:13:45.760 that's that's why really everything that we're that we're talking 0:13:45.800 --> 0:13:48.080 about has happened in the past like two decades. Yeah, 0:13:48.080 --> 0:13:51.800 it's pretty exciting stuff. So this one does have micro injection, 0:13:51.840 --> 0:13:54.679 does have some drawbacks. A big one is that you 0:13:54.720 --> 0:13:57.880 can't really conduct this UH in vivo, meaning within the 0:13:57.920 --> 0:14:01.400 body of a patient, because you can't you can't immobilize 0:14:01.440 --> 0:14:04.800 the cells. You can't really use the micro manipulator to 0:14:04.880 --> 0:14:07.520 inject something directly. All this kind of stuff tends to 0:14:07.520 --> 0:14:10.280 be done in a laboratory, in a special lab dish 0:14:10.440 --> 0:14:14.560 in the glass, right, Yep. You usually have a solution 0:14:14.679 --> 0:14:17.280 and you end up using a little pipette to immobilize 0:14:17.320 --> 0:14:20.640 the particular cell you want to UH to to manipulate. 0:14:20.720 --> 0:14:23.200 And now a lot of times they would use bacteria 0:14:23.320 --> 0:14:25.280 for this sort of thing, the idea being that the 0:14:25.280 --> 0:14:28.880 bacteria would then transmit the DNA to other cells. But 0:14:28.920 --> 0:14:31.600 we'll talk more about that and probably a future podcast. 0:14:31.680 --> 0:14:34.640 It's it's really kind of immaterial to this UH. There 0:14:34.720 --> 0:14:36.520 was another downside. It's that you have to do it 0:14:36.560 --> 0:14:40.440 to one cell at a time, which is not terribly efficient. Nope. 0:14:40.720 --> 0:14:45.560 And another UH drawback to this approach is that you 0:14:45.600 --> 0:14:48.160 have to be really careful exactly where you inject that DNA. 0:14:48.520 --> 0:14:50.840 They found out that if they injected the DNA into 0:14:50.840 --> 0:14:55.400 the cytoplasm, it was far less effective than if they 0:14:55.480 --> 0:14:58.720 injected it directly into the nucleus are possibly because the 0:14:59.040 --> 0:15:03.480 electro light salute in the cytoplasm was degrading the DNA 0:15:03.560 --> 0:15:06.040 before it could do anything useful. Right, So, you know, 0:15:06.200 --> 0:15:08.320 just like we said earlier that just because you put 0:15:08.320 --> 0:15:11.320 a new gene into a cell doesn't necessarily mean you're 0:15:11.320 --> 0:15:14.000 going to see that gene express itself or be effective. 0:15:14.400 --> 0:15:16.440 But they found that there was much more effective if 0:15:16.480 --> 0:15:19.560 they did inject it directly into the nucleus. Uh So, 0:15:19.600 --> 0:15:22.760 I mean it's it's certainly useful within the lab, but 0:15:23.040 --> 0:15:26.320 within real world quote unquote settings where you're trying to 0:15:26.360 --> 0:15:29.800 work on a patient directly, it's not. Now, if you 0:15:29.840 --> 0:15:34.240 can develop an approach a cure like within the lab 0:15:34.360 --> 0:15:36.680 within there and then implant those cells into the patient, 0:15:36.760 --> 0:15:39.720 that might be a way to go. But in general, 0:15:39.840 --> 0:15:43.200 this tends to be something we think about in terms 0:15:43.200 --> 0:15:45.960 of learning more about what's working and what's not working, 0:15:46.000 --> 0:15:51.800 as opposed to an approach um actual therapeutic treatment exactly. 0:15:52.160 --> 0:15:55.120 So that kind of leads me to another one hydro 0:15:55.320 --> 0:15:58.680 dynamic pressure, which again sounds you know, it's pretty much 0:15:58.720 --> 0:16:00.800 what sounds like, I didn't read that much about this one. 0:16:01.000 --> 0:16:02.880 Tell me, tell me what it is, all right. So 0:16:03.120 --> 0:16:06.840 they found out that if you take a relatively large 0:16:06.880 --> 0:16:10.600 amount of DNA material, and so for a human we're 0:16:10.600 --> 0:16:13.960 talking about actually just a few millilets, but that's relatively 0:16:14.520 --> 0:16:18.800 huge amounts of DNA, and then you take a needle 0:16:19.160 --> 0:16:21.840 and you insert the needle into a blood vessel inside 0:16:21.840 --> 0:16:25.960 that person, and then you inject that relatively large amount 0:16:25.960 --> 0:16:31.000 of DNA very quickly into that blood vessel. Then this 0:16:31.480 --> 0:16:37.080 the hydrodynamic pressure will cause cells along the blood vessels walls, 0:16:37.160 --> 0:16:41.760 the indothelium, that's the layer that the lining. Uh, it'll 0:16:41.800 --> 0:16:45.480 cause them to become more permeable, as well as the 0:16:45.680 --> 0:16:49.880 perenchyma cells or paranchema cells. Now these are like tissue cells. 0:16:50.160 --> 0:16:51.960 So this is like the cells of stuff that actually 0:16:52.000 --> 0:16:56.160 does work depending upon whatever you know, Oregan, you are targeting. 0:16:56.520 --> 0:16:59.480 So let's say it's the liver, all right. Well, they 0:16:59.520 --> 0:17:02.480 found that also increases their permeability. So if you have 0:17:02.560 --> 0:17:05.280 that DNA material and you inject it very quickly into 0:17:05.359 --> 0:17:08.719 the blood vessel, then you can end up having a 0:17:08.760 --> 0:17:12.040 lot more of that. Yeah, it ends up a lot 0:17:12.080 --> 0:17:15.520 more cells are going to accept that DNA and potentially 0:17:15.840 --> 0:17:20.240 you will start getting that gene expression, you're looking for. UM. 0:17:20.440 --> 0:17:23.239 Couple of other downsides to this. Now they've they've been 0:17:23.359 --> 0:17:26.600 using this with lab animals, not with people. All right, 0:17:26.680 --> 0:17:28.760 so this is not something that's been tested on people, 0:17:28.840 --> 0:17:31.360 but they've tested on mice and rats and it never 0:17:31.400 --> 0:17:34.400 got to the human stage because well, because they found 0:17:34.440 --> 0:17:38.440 out that sometimes this causes some cardial problems in the 0:17:38.520 --> 0:17:43.160 car cardial and respiratory. Yeah. Yeah, So when they would 0:17:43.200 --> 0:17:48.280 inject the vein along the tail of these critters, the 0:17:48.359 --> 0:17:50.439 mice and the rats. They did it with a mouse, 0:17:51.119 --> 0:17:54.080 the mouse would start to seem to have trouble breathing 0:17:54.200 --> 0:17:56.600 for a couple of minutes and then recover and everything 0:17:56.600 --> 0:18:00.760 seems to be okay. Rats could sometimes stop breathe entirely. 0:18:00.840 --> 0:18:04.239 And they found that by massaging the abdomen's they could, uh, 0:18:05.119 --> 0:18:07.720 they could they could end up getting the rats to 0:18:08.160 --> 0:18:11.040 you know, kind of start breathing again. But they discovered 0:18:11.080 --> 0:18:14.280 that using this approach put a lot of pressure on 0:18:14.359 --> 0:18:19.520 the animals uh cardiovascular system, and that it would cause 0:18:20.040 --> 0:18:24.440 everything from labored breathing to uh, irregular heartbeat. Uh. They 0:18:24.440 --> 0:18:28.439 also discovered that the animals livers were expanding, sometimes up 0:18:28.440 --> 0:18:33.359 to two of their original size. I guess that weirdly 0:18:33.840 --> 0:18:39.600 makes sense. I mean, in terms of of liquid essentially, 0:18:39.640 --> 0:18:43.000 you're essentially overloading the system, right, you know. And uh, 0:18:43.080 --> 0:18:47.920 and so none of these um issues were permanent unless, 0:18:47.920 --> 0:18:50.680 of course, the animal did not survive that initial reaction. 0:18:51.000 --> 0:18:55.399 But the animals survived, then their various systems would return 0:18:55.400 --> 0:18:58.159 to normal, Like they're breathing and heartbeat would return to 0:18:58.160 --> 0:19:00.920 noble normal within a couple of minutes. Uh. Their liver 0:19:01.080 --> 0:19:04.000 would go back to being normal sized within about half 0:19:04.000 --> 0:19:06.760 an hour, and some of their other systems would take 0:19:07.040 --> 0:19:09.200 up to two days or a day and a half 0:19:09.280 --> 0:19:12.800 or so to get back to normal, but they started to. 0:19:12.960 --> 0:19:16.720 They also would very frequently start expressing whatever the gene was, 0:19:16.800 --> 0:19:20.480 so in front that side, it was a success. So 0:19:20.600 --> 0:19:23.399 all of these so far have been interesting, UM, but 0:19:23.560 --> 0:19:27.880 perhaps not the best way to actually treat human patients. UM. 0:19:27.920 --> 0:19:30.680 We've got a couple more to talk about. But first, Yeah, 0:19:30.760 --> 0:19:34.959 let's take a quick break to thank our sponsor. All right, 0:19:35.280 --> 0:19:37.639 we're back. So we've talked a little bit about a 0:19:37.640 --> 0:19:41.280 couple of different approaches that may we may never see 0:19:41.400 --> 0:19:44.239 used as far as human applications go. I mean, there 0:19:44.240 --> 0:19:46.880 are there's work in hydro dynamic to try and make 0:19:46.920 --> 0:19:49.920 it a something that we could use with human patients. 0:19:50.000 --> 0:19:52.960 But anything that would put that much stress on your 0:19:53.000 --> 0:19:56.240 cardiovascular system for someone who's already trying to undergo medical 0:19:56.280 --> 0:19:59.119 treatment is definitely not going to be the first choice, 0:19:59.240 --> 0:20:03.160 right right, No, So let's talk about um, well, let's 0:20:03.160 --> 0:20:06.040 talk about zapping cells. I was just thinking, you know, 0:20:06.080 --> 0:20:08.800 if we can't if we can't really stab a needle 0:20:08.840 --> 0:20:11.800 into a cell, if it's inside a person, and if 0:20:11.800 --> 0:20:15.240 we can't really expect to jab a bunch of liquid 0:20:15.280 --> 0:20:17.480 into a blood vessel and possibly cause a heart attack, 0:20:17.880 --> 0:20:21.840 maybe we could just shock the heck out of them. Yeah, 0:20:21.840 --> 0:20:25.600 that's clearly the next logical step. Um no, no, but 0:20:25.640 --> 0:20:28.040 actually it does make a lot of sense. Yeah, because 0:20:28.080 --> 0:20:30.080 I mean, we've got those membranes. They're not going to 0:20:30.240 --> 0:20:32.760 let that DNA just go right on in. So how 0:20:32.760 --> 0:20:35.600 do we convince it that we need to get this 0:20:35.720 --> 0:20:40.000 DNA from outside the membrane inside the membrane? Right? Okay? 0:20:40.040 --> 0:20:43.320 And one of one of the really cool and important 0:20:43.520 --> 0:20:45.600 facts about cells that that you that you need to 0:20:45.600 --> 0:20:48.399 realize here is it. Cell membranes are basically insulators that 0:20:48.440 --> 0:20:52.720 are separating two charged regions, the the electrolyte or ionized 0:20:52.800 --> 0:20:56.359 solution inside the cell and the electrolyte solution outside the cell. 0:20:56.760 --> 0:21:00.240 So so they can they can act as capacitors. They 0:21:00.320 --> 0:21:03.159 have they have capacitance. They do have capacitance, and if 0:21:03.200 --> 0:21:07.159 you overload that capacitance, things go wonky. Stuff happens, right, So, 0:21:07.200 --> 0:21:10.600 if you do it at the right amount of of charge, 0:21:10.960 --> 0:21:13.960 then it ends up creating these pores, these these temporary 0:21:14.000 --> 0:21:16.520 pores p o r e s. So you get these 0:21:16.520 --> 0:21:20.000 little holes that form in the membrane, and it's temporary. 0:21:20.040 --> 0:21:24.080 They will actually heal back up actually actually just a 0:21:24.080 --> 0:21:27.120 few minutes. Yeah, within a few minutes after zapping them, 0:21:27.119 --> 0:21:29.520 assuming that you're zapping them with the right amount of electricity, 0:21:29.520 --> 0:21:32.800 because that's really important. So if you if you do 0:21:32.840 --> 0:21:36.280 it at a low enough UH or well really it's 0:21:36.520 --> 0:21:43.080 short high voltage zapps um very low amperage, and so 0:21:43.200 --> 0:21:46.600 you're just doing these tiny little controlled zapps to the 0:21:46.600 --> 0:21:49.120 the cell membrane, these holes will open up and then 0:21:49.119 --> 0:21:52.199 within a few minutes after you've applied it, they'll close 0:21:52.240 --> 0:21:55.520 back up again. So while they're open, then you can 0:21:55.560 --> 0:21:58.879 coerce d NA to go in because DNA's are charged 0:21:58.960 --> 0:22:02.080 particle and right so so especially since you've just changed 0:22:02.119 --> 0:22:05.480 the capacitance of that membrane, it makes it easier for 0:22:05.560 --> 0:22:08.240 stuff to slip through when it's also charged. So you 0:22:08.320 --> 0:22:10.679 just you kind of you kind of corral it in 0:22:10.960 --> 0:22:13.600 so you're you're you're driving the DNA and it's not 0:22:13.640 --> 0:22:16.520 like it's just gonna you know, oh yeah, zip. You 0:22:16.520 --> 0:22:20.600 actually have to guide it yourself, yourself being if you're 0:22:20.600 --> 0:22:22.719 a scientist or doctor who's doing this sort of thing, 0:22:22.720 --> 0:22:26.200 obviously probably not like no, no, no, no, probably would 0:22:26.200 --> 0:22:28.639 not do this. I think Noel does want to stab 0:22:28.720 --> 0:22:33.720 me with stuff, but probably not in a medicinal way. Anyway. 0:22:33.800 --> 0:22:38.240 This is called electroporation. Obviously you're you've got the electro 0:22:38.359 --> 0:22:41.040 and the pooration. You've got the making the poor. So 0:22:41.080 --> 0:22:43.040