go, I think this is going to work. Okay, so remember, we're getting into this faith in
a novel virus, and they have faith in a novel biology, it's actually morphed into this faith
in a novel virology, a virology, excuse me, that revolves around a gain of function RNA
virus, it involves around particular, particular pieces of that RNA that made it extra special,
it involves transfection working, but not working for being rushed, being adulterated
or having chosen the wrong protein. That wrong protein is the spike protein that from
very early on in 2020, here in the first year already, was being lambasted by many different
people in the narrative and in the media about being a gain of function protein that the
protein itself was the evidence. And then later the protein is a toxin later, the protein
was causing amyloidosis, prion disease, etc. And so this whole, this whole mythology,
this immuno mythology that they have, they have, they have seeded around the novel virus,
has gotten quite, quite expansive. And the part that we're working on here with these
two streams, the one that I just ended in the one I'm starting now, is understanding
protein folding in the context of prion disease, so that we can get an idea of where they
have exaggerated, where they have simplified, and how we can, how we can bring this back
into a really reasonable focus where the sacred aspects of biology, the irreducible
complexity of biology is met with a certain reverence that right now it's, it just is
not given. So what we're really working with here is a faith in a novel biology, it's
a mythology that covers for the expected damage as we transition into population wide testing
of transfection technology. So they know that amyloidosis and prion disease and protein
misfolding and this kind of damage are going to occur over time if they continue to transfect
old people and young people alike. And they, they expect it to go up, they expect it to
show up from already the, the people that have been transfected multiple times. And
I believe this because it was also included in the spars pandemic narrative on page 48,
you can find a little talk about how three years after the rollout of the, of the vaccine
for spars, that a bunch of people develop some kind of, of, of crowds felt yock of disease
and it was blamed on the, on the vaccine and a lot of people were upset. And it started
to undermine the way that people felt about the public health system. It's in their little
Rockefeller tabletop exercise called the spars pandemic, you can find it. And so without
a doubt, what they learned from that tabletop exercise and others was that if they wanted
to cover up for this, the easiest way to do it would be to see a lot of these potential
worst case scenarios into the gain of function story in the beginning of the pandemic. So
that by the time those things manifested, it would be too lost in that narrative of lab
leak or natural virus. And the whole acceptance of a circulating novel pathogen would have
already happened years earlier. And that's where we are right now. That's why, that's
why we need to watch this.
Guess it was me that was, it was clipping here. Somehow I have everything. Oh, I see it
now. Sorry, I got it. I got it. I got it. It was a knob, a knob, a very knob that had
been brushed by a looks like it got pulled by the headphone wire there. Now I see it. Now
I know why my my my my levels were. So there, there was the little morbid title that I was
going to use proteins and protein folding with a dead lady from MIT. That's this one. I'm
going to get rid of this. And I'm going to bring up a new window from my word file here, part B.
That would be this one. So we got the idea that maybe we could use those these cells as
I'm sort of living test tube. And the reason why we would want to do that is there's no
organism. So this is starting in the middle. I got to go to the front. Sorry about that. Here
we go. So here we go. I'm going to put it at 1.5 speed. Hope you can handle it. I think that
should be normal. Now we're used to it.
Remember the Howard Hughes Medical Institute and I work at the Whitehead Institute at MIT. I work
on a variety of different protein folding problems. And in my last lecture, I told you a video
brought introduction to the problem for how it manifested. I think it's a little bit
how it manifested in infectious diseases and more broadly how it is used by cancers to drive
them in the living state. In this lecture, I'd like to tell you about a different aspect
of protein pathology, another equally devastating aspect of protein folding and pathology, the
neurodegenerative diseases, because all of these diseases are diseases of protein and
swollen. This is a extremely vivid demonstration of the difference between the brain of a normal
adult, a pine autopsy, versus adult who died of Alzheimer's disease. It's obviously devastating
disease. And this is why the people who have these diseases lose their memory, lose control
of functions. Okay, sorry. Diseases are really terrible.
Now, this is a graph of what's happened to human longevity over the last couple hundred years.
And it's really, I think this is the red and the black are just two different calculations.
It's not so easy in the going back to the older days to calculate when exactly how old people
lived on average. But these two very different ways of doing it came out with the same answer.
And you can see that there's been the steady march of progress and it's just been amazing.
Wait, where's the drop off for is World War I then going to be considered the flu? I thought
the flu showed up on that one graph all by itself. It was just the flu not World War I or
anything like it was just the flu. Remember? Holy cow. Now she says it's World War. So that's cool.
This has been I think one of the glories of mankind to be able to do this and alter their
own average lifespan. And it's been due to many different factors due to changes in public health,
cleaner drinking water, due to refrigeration and preservation of food and cooking. It's due to
the development of antibiotics, the development of vaccines, the development of anesthesia. So
you could do surgery on people and correct illnesses that way. So anyway, this wonderful
steady, steady progress of mankind is unfortunately in some ways of thinking about it a road to ruin
because as we are curing these other diseases, as we're living longer and longer lives,
we are finding that the incidents of neurodegenerative diseases are going out. These diseases used to
be practically unheard of 100 years ago. Now there's a very large fraction of people around the world
that are suffering from these diseases. And as we extend lifespan, it's getting worse and worse.
There are 5 million Americans suffering from Alzheimer's disease alone. And the same increase
in disease is occurring for all of the neurodegenerative diseases across our globe. So unfortunately,
with respect to neurodegeneration and it being a road to ruin, this is why I say a road to ruin,
we're headed for neurodegeneration and right now there's no exit. We do not have a single therapy
that really fixes these problems. So these are some of the common and uncommon neurodegenerative
diseases you might have heard about Alzheimer's disease and Parkinson's disease, frontal temporal
dementia, Huntington's ALS and Croissphal Yacob disease. And you can see these brown blobs
inside of these cells. And those brown blobs are aggravated proteins, like those aggregates of
Friday I showed you earlier. And as I said, there are all these neurodegenerative diseases
are protein folding diseases and there's not a single therapeutic strategy that cures the
underlying protein pathology. We have some things that address some symptoms in some of these diseases,
but for the most part, we're pretty helpless against them. So I've been working on protein
folding for a long time and I've worked on a lot of different organisms. And the one thing
that my studies over the years have taught me is that this problem, as I mentioned earlier,
is common to all organisms on earth. And so we got the kind of crazy idea that considering the
eukaryotic tree of life, you see, plants, animals and fungi actually split from each other not that
long ago in terms of evolution. So we thought we might be able to take advantage of this similarity
and to study some of these really difficult, really complicated diseases. Yes, we will not be able
to study many different aspects of protein folding neurodegenerative disease in a simpler
organism. But if we could study some aspects of the precipitating, initiating protein pathology,
the cellular pathology, not the complexity of the disease as a whole, but just the initiating,
precipitating pathology from those proteins in a simple organism, we might be able to move
much more quickly than we would if we had that we can find solely to working on these more complex
organisms. So as I mentioned, one of the things we have in common with yeast is a wide variety
of systems for controlling the protein folding problem. So we have chaperone proteins which
interact with highly reactive proteins that are not quite finished folding and prevent just like
human chaperones, prevent them. Their charges from interacting inappropriately with other partners
and until they're ready and mature, protein chaperones do the same thing. But we also have
protein modeling factors, things that can rest those protein aggregates when they start to appear
apart. We have osmolites, we have things called leproteosome, which degrade proteins that are
that are not properly folded, ubiquitin, ubiquitin ligases, and that entire system is just completely
conserved from yeast to human cells. It's not just that. Lipid biology is actually quite highly
conserved. There certainly are differences of lipid biology at least in human cells, but for
example cholesterol, yeast use a very closely related lipid called regastrol for exactly the
same reason that we use cholesterol to control the fluidity of membranes and to control the movement
and density of proteins within those membranes. And they move packages of membrane-bounded proteins
around the cell in very highly orchestrated ways, really the same way that a nerve cell will move
dopamine around. The yeast cells will move things like mating factors around.
So she says that we move dopamine around in little vesicles. That's all we do is
neurotransmitters around in little vesicles. That's really sad because we know in the brain there are
at least vesicles of RNA that are released of the arc protein, which cause local actin skeleton
remodeling to be possible in neighboring cells that weren't necessarily activated by the same
genetic or neuronal signal that the postsynaptic neuron was. And so the postsynaptic neuron can
release virus-like particles that contain the mRNA of the arc gene cause arc protein to be
expressed locally at that synapse and cause remodeling of that local arc protein cytoskeleton
in that neuron that was never activated by the synapse or the signal that came into the postsynaptic
neuron. And that is just the tip of the iceberg. I'm quite certain of it. So it's really funny,
almost to the point of being a little bit... Why did you say that? If you say that like we
move dopamine around these yeast proteins move all kinds of stuff around in their vesicles.
We know that there's extracellular signaling between tissue using extracellular vesicles
called exosomes. I'm sure she knows it too. She's got to know it. And if she doesn't, it's because
of the intense compartmentalization of biology. Sosomes and peroxosomes, these are very complex
organelles that are involved in doing very complicated functions. Some of them are involved in degrading
proteins. Some of them are involved in a wide variety of metabolic actions that have to be
segregated from the normal cytoplasm. These cells have both of those. They have autophagy. This
is a process by which the cell actually directs its degradation and eating machinery to eat up
protein aggregates and get rid of them. Apoposis, a programmed form of cell death.
Cell cycle, very complexly regulated cell cycle, regulated very, very differently bacteria,
but in yeast and humans regulated in very much the same way. And in fact, studies of that cell
cycle work extremely important for our understanding of cancer. And why cancer cells start to replicate
uncontrollably, study them in yeast to provide its key insights. We have mitochondria, the
energy factory of the cells, and mitochondria do amazing things in yeast and human cells.
But they also are a place where reactive oxygen species are generated and can do a great deal
of damage. And then there's a whole variety of signal transaction pathways. Again,
these key pathways that control growth and development in us, but control responses to the
environment, responses to other cells, and responses to internal and external stresses,
those same signaling pathways have been controlled, have been preserved rather in yeast and higher
eukaryotes. So, calcinarism, example, map kinases, G-coupled protein receptors,
all of these were first developed long ago in eukaryotic life, and greatly greatly elaborated
in us. We have many, many more G-coupled receptors than a yeast cell has, for example.
But the basic machinery and the basic concepts and the basic ways in which those signaling pathways
drive processes inside the cell are similar. So, we got the idea that maybe we could
use those yeast cells as our living test tube, and the reason why we want to do that is there's
no organism on Earth that we can manipulate and get to tell us its secrets better than yeast.
It has an absolutely unrivaled toolkit, and it really derives from brewers back about 150 years
ago wanting to make better beer, and wanting to understand that organism and how to manipulate it,
and it's taken off from there, and it's just amazing. Massive, massive numbers of people have
been building and developing technologies that allow us to knock out every gene in the genome,
or overexpress every gene in the genome, make point mutants wherever we want in the genome,
and so that's just something we can't do in any other organism at this level today.
So, here's how we set things up. We have yeast cells that are growing on, in the top row there,
they're growing on glucose medium. In the bottom portion of the panel, they're growing on lactose
medium. We have a gene that will turn on whenever we give the cells lactose, and so we make a
recombinant form of that gene that now, well, instead of making the proteins that these cells use for
lactose utilization, they make different proteins that misfold in human diseases, like alpha-synuclein,
A-beta, TVP-43, and TIN-TIN-FUS. And you can see that we've built, for synuclein here, we've
shown you all three different strains that are expressed in a protein at different levels,
and are exhibiting different levels of toxicity, just by the fact that they can't grow very well,
and we've then done that with all of those different disease proteins, and we've matched them so that
they have the same level of toxicity. So, same level of toxicity from different proteins,
what I said is just some non-specific protein aggregation mess. It turns out that it's not,
but when those proteins misfold inside of the yeast cell, they go into the cell, they interact
with the same kinds of highly conserved constituents that they interact with in a neuron, and they do
bad things in a very specific way. So, here's an example of a phenotype that glob over there
is protein nitration, and it's happening all of the cells at the same level of toxicity,
the nitration damage is happening really only in the cells that are expressing
alpha-synuclein. That's really interesting, because in the human diseases that are known to be caused
by the misfolding of alpha-synuclein, and that is Parkinson's disease, multiple systems atrophy,
Lewy body, dementia, and nerve-brain iron accumulation, they too show very high levels of very specific
protein aggregates with nitration. So, very unique and very specific cellular pathologies directly
related to the human disease. So, here's our cells, we've got this gene that we can turn on with
galactose, anaerobic galactose, and we've hooked it up to GFP, just so that we could see what was
happening to it in the cells, as they were either healthy or a guy. And when we had just one or two
copies of the protein in the cells, they would find, and the protein went out to the membrane,
which is where it should belong. And if we had more, one extra copy, we started seeing things
going along, and then if we had two extra copies, it went even worse, this does not look good,
they said protein conglomerates here in aggregation, type some type of aggregation, and then those
cells grow fine, those cells grow slowly, and those cells die. Very, very strong dosage difference,
and what's really interesting about that, is that's true in man as well.
So, it's really important for you to understand that there is one gigantic caveat here, which I
find almost disturbing. Green fluorescent protein is a massive protein, native to jellyfish that
glows in the dark green. If you attach that massive protein to alpha-synuclein, I would be
willing to bet that that cartoon up there is wrong, the GFP is a lot bigger than alpha-synuclein.
Could be wrong, you look it up yourself and find out if I'm right or wrong, or whether that cartoon
is right or wrong, but the point is, is that GFP is not small, and overexpressing GFP in any cell
line, or in any mammal tissue will result in cell death. Because that level of GFP is toxic,
if you can see it like this, it's already a lot of molecules, a lot of molecules of GFP in order
to see that signal. If it starts to make these kinds of, these kinds of, uh, punctate sort of
constructs of, of, of the cell is, is putting this GFP into vesicles to get rid of it, to keep it,
to keep it compartmentalized, so it's already beyond toxic levels. And she is very conveniently
ignoring that fact, because there are no controls here with, with just GFP, right, to show what
GFP toxicity looks like, but I can guarantee you, I can tell you from experience that GFP
toxicity is real. Over expression of GFP causes toxicity is very, very real, and that is toxic
levels already in the middle. And anybody that's used GFP to label neurons in a mouse brain could
tell you that anybody that's used GFP as a label for anything can tell you that because if you get
a good good signal, you also get dead cells. It's extraordinary. And this is the kind of science
that passes for knowledge creation at this time. This is how we got here. This is, you know, how
many years ago is this eight? Just the basic principle of this is silly because the GFP is not
just a glow in the dark tag. It's a 10,000. It's a huge thing. Let's just, I'm just going to look
it up because I don't know. Green, fluorescent protein, and there will probably be a Wikipedia
page right there. Green fluorescent protein has a wavelength blah, blah, blah. Natural protein is
238 amino acids. It's 27 kilodultans. And then let's look up alpha synuclein.
140 amino acids. So what is it bigger?
238. No, it's bigger. 140 by 238. So it's roughly twice the size of it, you see? So this is not the
right, this is not the right cartoon. It's a much bigger protein. And she is pretending that that
much bigger protein has no effect on whether the cells are healthy or not. It's all the alpha
synuclein. That's it. That's impressive.
Things that have just one extra copy of the wild type of synuclein protein will get early onset
Parkinson's disease. And if they have two extra copies, they'll get even earlier, more virulent
form of the disease. So this unusual, I mean, ask yourself, why does it need to be tied to GFP?
If you know that alpha synuclein is being expressed by the cells, why can't you stain for it? Why do
you need to tie it to GFP? When GFP is a gigantic toxin, a gigantic toxic protein at high fluorescent
levels, it's extraordinary. Extreme sensitivity to exactly how much protein you're making was
certainly, was certainly reminiscent of what was happening in man. So how can we get a better idea
of what's going on here if there's anything really deeper involved? Well, we do something
called screen first, you could do a control for the GFP. Meaning we screen every gene in the genome
for what makes cells better or worse. We can take with these, we have libraries,
every gene in the genome, we can turn them up or turn them down and see how that changes
the disease manifestation. And in these cells that have the four copies where they're just
playing frankly dying of the disease, we can screen for chemical compounds that might rescue them.
And studying those compounds might tell us something about the disease mythology.
So I love the fact that she uses mite, but it's exactly how science is done. You declare
a rough shot experiment, a suitable model for a disease, and then you go for it.
That's what she did just there, right? She said, this is a pretty suitable model for disease.
Why don't we use the four copy version for a model of disease and we'll screen compounds
on it? Holy cow, that'll pay for at least two postdocs in five years of my lab.
Compounds that might rescue them. And studying those compounds might tell us something about
the disease mythology. So screening is a lot like panning for gold. You go through a whole
lot of stuff and you look through it, you look through it and you look through it and you find
nothing for a while, and then all of a sudden you get these and then get some gold. And you get,
so out of the 6,000 genes in the yeast genome that we studied, only about 60 or 70 of them in
our initial studies seem to matter with respect to alpha-snooplane. And the genes that we got out
of our alpha-snooplane screens were completely different than the genes we got out of our
beta screens and completely different than the ones we got of our proteins in the screen.
And they told us something about the biology. Because for example, the largest class of genes
we got were genes that were involved in busgal trafficking, moving those numbering bounded,
proteinaceous compartments around the cell. And so when I showed you these protein conglomerations
or these aggregated forms of protein in this cellular model of the alpha-snooplane pathology,
it turns out that when we got that result, the genes that saved the yeast cells from that pathology
were genes that were involved in moving little vesicles around. We thought, well,
gee, I wonder if those things actually have something to do with vesicle trafficking. And so
when we look at the level of the electron microscope, which allows a much, much higher
resolution of the cell, you can see that, yes, these little vesicles that are packed with proteins
depending on how much the alpha-snooplane we're expressing, we get more and more of these protein
aggregates. And then we did something called immuno-electron microscopy.
This is absolutely terrible, right? You know, if this is the same cell model that she's looking
at under the microscope here, under the electron microscope, she has no idea whether this is from
the alpha-synuclein or whether it's from the overexpression of the GFP, or whether the
combination of both would do it. I just can't, I can't really fathom how bad this is. I got to
believe that these papers have some controls or something in them, but I'm scared that they don't.
We attached a label to an antibody against the alpha-synuclein and against a protein involved
in vesicle trafficking, and we found that they were there together. So these blobs, these green
blobs here, are actually blobs, not just of aggregated cineuclin, but aggregated cineuclin
enmeshed in vesicles that are not moving around the cell and getting to the places they're supposed
to be. And when that happens in a nerve cell, it's really disastrous because that's one of the
major ways in which a nerve cells communicate with each other. That's so good for a yeast cell
either. Anyway, this finding that alpha-synuclein blocks vesicle trafficking has not been corroborated
by many other laboratories. And to cut a long story short and move on to the very final stage of
this talk, we found that there were parallel effects, we moved back and forth between these
two neurons, and we found that there were parallel effects. We moved back and forth between yeast
and neurons, and you've got to be very careful when biologists are claiming to be able to do things
like that. Not just vesicle trafficking, but bursts of reactive nitrogen species, as I showed you in
that protein block, mitochondrial dysfunction, and perturbations in middle line homeostasis. So
at least at this early, very simple cellular level, there's a lot of similarities there.
But we really needed to be able to show that the genes we found in yeast, and the genes that
saved the yeast cells, those same genes would matter to a neuron. So we actually looked at,
in a couple of different systems initially, one was this wonderful nematode system,
it was a worm, it's a simple little worm, but it's got lots of different kinds of neurons,
and in fact it's got the same kind of neurons, dopaminergic neurons, that are adversely affected
in Parkinson's disease. And we could actually peer through, wire up those cells to express
alpha-synuclein, and mire them up so that they were green, they glow green, we could actually
study them in a living worm, and we could see that when the worms were expressing alpha-synuclein
in those cells, you can see how some of them are disappearing over there. It's a true neurodegenerative
model in the nematode. And our genes that rescue the yeast cells also rescue that nematode.
And the same thing happened when we took neurons from rat brains, the midbrain region of the rat,
which is the corresponding region that's affected Parkinson's in humans. So that was pretty encouraging.
Next thing we did was to screen a chemical lab. Of course there she said that she's taking embryonic
rat neurons from a particular brain region, and then she's culturing them and using them as a model as
well. Just be clear, this one looks like it has a GFP-only stain, so this one looks like it's GFP
only, this one looks like it might be, then alpha-synuclein, I got to get this alpha-synuclein
plus GFP. I mean, it seems like that's a better experiment than the one she showed us, at least
there's some evidence there was a control there. So you could assume that maybe there was a control
back there where I was losing my mind, but I'm not sure. I don't know. I hope so.
So we're doing here is taking a human protein, expressing it in C. elegans, and then having those
neurons die. Is that really surprising? Alpha-synuclein would be a protein that would be foreign
technically, right? However, they got it in there, however they put it in there and be interesting to see
to think about, what did she say she did with this? Six hits from yeast screen validated in both
nematode and neuron models, so the midbrain region of the rat. Which is the corresponding region that's
affected Parkinson's in humans. So they want to be used to rescue the yeast cell, also rescued
that nematode. And the same thing happened when we took neurons from rat brains in the brain region
of the rat, which is left. And the same thing happened. So supposedly they expressed alpha-synuclein,
and then those those, but overexpression of protein doesn't necessarily is not surprisingly
toxic. That's normally toxic. So I'm still curious as to where this is going. A corresponding
region that's affected Parkinson's in humans. So that was pretty encouraging. Next thing we do
is to screen a chemical library. And this again is something that is so much easier to do in yeast.
We asked whether we could find compounds that would fix more than one problem. I told you there
lots of different things going on. There's a cascade of pathology that gets kicked off by those
misfolded proteins. And can we find one that compounds that can fix more than one of those
problems? So and the next question was can we use yeast genetics to find a target? So why would
this matter? Well, we could screen through. Can we use yeast genetics to find the target? So you
see what she's doing here. She's developing the same kind of screening techniques across genome
libraries that would eventually be employed by the actual project on humans. He just
need the computing power in the data facility. And this is at MIT Whitehead. It's at the same
institute that developed all the stuff for the human genome project. It's at the same institute
from which the patents that that Kevin McCurnan built his his many companies from came from.
And so we're talking about all the same basic mythology, which is designed to make you think that
on the basis of studying disease and public health that that they're they're developing
techniques to help people when in reality they are developing techniques, which will ultimately
be used to mine the human population for as much medical and genomic data as possible.
And of course, they have to start with something simple that has all the basic parts of our
cellular physiology like yeast. And if they can do it in a single-celled organism like yeast,
then they can eventually expand to a multicellular organism and eventually to us. That's what this
is all about. Did it that screen through 500,000 chemical compounds asking for which ones were
able to rescue these cells? That kind of a screen which took us several months would take, I don't
know, maybe a hundred thousand years if you were using a mouse. And also probably, I don't know,
billions and billions of dollars. We did it much more cheaply, much more easily, much more rapidly
in these cells. And of course, you heard Ray Kurzweiler say that we're not going to do this in mice
or in people. We'll do it in synthetic computerized people, synthetic biology and in silico biology.
The other reason why it mattered was that these cells offered, as I mentioned earlier, they are
unparalleled genetics. And so we're actually able to take advantage of that genetics to figure out
what those compounds were doing to save the cell and then go back into neurons and ask
whether those same compounds would work in the neurons and would those compounds fix the same
pathologies that are taking place in the neurons. And it worked. So we screened 500, 50,000 compounds.
Simply asked for restoring growth. We've got a lot of genes, don't know which one is the right one
to try to go after. We just looked for something that would have stored growth. And we've only
dissected a few of these compounds so far, but they merely raid vascular trafficking defects,
they merely rate mitochondrial defects, and they work the ones we've tested in nematode,
react, and human neurons. So the final piece of the story is to turn towards human
IPS cells made from patients that have one of these diseases. This has been one of the most
exciting aspects of revolutionary aspects of biology in terms of being able to devise better
treatments for patients. And so this is interesting because this is one of the technologies that I
think when I was going into neuroscience, what I thought was kind of cool was that they're going
to do is they're going to take cells from the patient, they're going to induce them to form
the tissue that you want to study in the laboratory that you think would have in
genetic similarity to the patient in question. And then you could do your screening on the
preparation that was made from the patient's cells, but never need any patient sample other
than that. I thought that was kind of a cool idea in theory.
Skin cells from a patient can actually de-differentiate those skin cells into an embryonic surge state
and then re-differentiate them into neurons. And another amazing technology that's been developed
recently by many other investigators has been the ability to surgically genetically edit
those cells such that you have corrected just the mutation that's responsible for that person's
disease. And so you have absolutely genetically identical cell types here. The only difference
between them is the difference that causes the disease. Well, there you go. That's what CRISPR
now can do or they say can do with a lot of off-target side effects. But even Jessica Rose says the
plan is to use CRISPR for personalized medicine. She announced it on our last
substack as well as trying to speculate as to whether they're not already CRISPR-ed you in the
shot. But definitely they will in the future. It was like a big announcement. But the idea that
we should resist or that there was something to be stopped was not in that substack. It was actually
she was sure that CRISPR was part of the plan in the future of personalized medicine and genetic
modification of humans. Well, that's really great. And here it is already 10 years ago. They're
talking about this stuff and they knew that this was the plan. They knew this was the way they were
going to go and they slow walked us there. And I think that this MIT professor was probably part
of it. They have to get you to believe that protein folding is a, you know, criticality.
And it's a criticality upon which all of these pathologies depend. And then they get you to believe
that because that's what they're focused on. Everything they look at has a play. And so in this
case protein folding has that role. And so then you can ask whether or not you have any pathologies
that are different between them and whether any of the things you've discovered earlier
work against those pathologies except the cells looked pretty much identical. So how do we figure
out what pathologies might be happening? Because after all these pathologies only manifest themselves
in terms of human disease, even if people have these terrible mutations, they only manifest at
the ages of 40, 50, 60. Yeah, so if you get all of academic biology to work on IPS cells for 10
or 15 years, then at the same time you get all of biology to develop very, very, very,
how do you say it homogenized and optimized techniques for using and leveraging IPS cell
technologies. And that could very well include using them as medical technologies, medical
interventions, even longevity, this kind of thing. And you don't have to have any academic
biologists be aware of that. You just have to have the ongoing infrastructure to make sure that
the remnants from all the hospitals and all the world that can produce IPS cells are always available
and are always flowing and are always coordinated and are always there. And that's the way it is.
None of that material goes to waste. It's way too valuable. And because it's not going to be
there forever, we're not always going to have 350 million people in America and 100 million
people needing a hospital every year. We're not going to have that forever. They don't plan to
have that forever. So they need to gather all this data now. Seven years old. But we had the ability
to go back to these cells and remember what we had learned from these cells and looked for
those same pathologies arising in these cells long before they started to die. So long before
this has started to die, we saw problems in the same problem in the vascular trafficking,
the same problems in nitrosative damage, nitroic mitral stress. And when, and they did not happen
in our Newton corrected cells, so that made us allow us to know that yes, not only were those
pathologies happening, but those pathologies were due to the mutation that was causing that person's
disease. And then we went back and we asked whether or not our compounds could rescue those cells
and they could, at least the first few that we've tried have. And we then used yeast genetics,
as I mentioned, a lot of complicated experiments I won't take you all through. But we used yeast
genetics to figure out what the target of the compound was. And lo and behold, it turned out
to be a very, very highly conserved ubiquitin ligase. You can see this is a cartoon of the
different domains of the ubiquitin ligase in the yeast cell known as RSP5 and in human cells known
as NED4. And basically every domain is conserved. And this is really an interesting kind of protein
defined because the ubiquitin ligases are very large complex family in humans. There are about
700 of them in humans. There are about 300 of them in yeast. And they've been very,
very difficult for pharmaceutical companies to target when they take the protein out of the cell
and try to do what they have done traditionally over the years to try to find chemical compounds
that will alter the function of the protein in a purified system. And that is because the biology
of these proteins really only manifests within the context of a living cell where all the proteins
are very, very close together and crowded and moving around and changing their conformational
states. That's where you see that this protein particularly matters. And this protein is very
complicated when it would have been impossible to find without the kinds of simple chemical genetic
methods that we use. So it really demonstrates I think pretty strongly that the power of phenotypic
screens, if you're looking for compounds that really have very special properties, properties that can
correct the disease pathology and the power of chemical genetics to figure out how those targets
work. Now this is really only the beginning whether this will ever turn out to be a therapy or not,
we don't know. But what it has done is showed us that this ubiquitin ligase plays a very key
central role in the way that pathology, that cellular pathology is manifested in lots of different
ways. And so it's certainly a useful tool. We're finding more and more of the genes that we found
in these cells are useful tools in understanding the biology. And we hope one day maybe this will
be a way of finding therapeutic compounds. But what I do believe is that we need these diseases
are very, very difficult and we need to try every trick in the book. And so something as unconventional
as this, maybe it could provide one key. So we're pursuing the same sort of strategy with
Alzheimer's and other neurodegenerative diseases. Many other people are pursuing other very
strategies. But the reason I concentrated on showing you what happens with one particular
protein was that, remember that idea that maybe you could alter this HEAC response and
and soup up a HEAC response in these neurodegenerative diseases and take care of the protein folding
problem? Well, our work with cancer told us that might not be a good idea. It might make those
the brain cells much more susceptible to cancer. So that's why we're going after individual proteins
in these diseases. So the first and final thing I want to say is that this work is all thanks to
the extraordinary group of people in my laboratory who've been working with their whole hearts and
souls over the last. So I wonder if you can already notice that one of the shocking things for me
is that we've never really got to prions. In this two part lecture, we never really got to prions
and what prions are and what prion misfolding is. We never got to aggregation or this kind of
thing, which is also thought to happen with some of these proteins. So it's really curious to me
because over expression of alpha synuclein being some kind of toxic thing is not very surprising to
me. Any over expression of a protein could potentially be toxic. I just find a lot of this
stuff to be. I'm sure that there's some good science there. And I'm also sure that there's a
lot of exaggeration, especially with regard to prions. So I think tomorrow we're going to take one
last look at her and prions. And then we're going to move on to the main literature of the Nobel
Prize by Stanley Pruser. And we're going to do our little short course on prions and protein
folding and where we are then and where we are now. And see why it is, for example, that Google
Fold hasn't told us how prions work yet, even though Google Fold has supposedly figured out how
protein folding works or it's really, really good. It's quite shocking. And when you see it
from stepping back, it's really very easy to see how overextended they are. And you know,
that's the way it is. They have told us lots of stories about what artificial general intelligence
is going to do. And what it really is is a story about that they tell each other in private meetings.
It's a story about how they have to use this resource while it's available. And as we ramp down
the reproductive rate of our global population and dissolve national borders,
we want to be sure that we collect the data from all of these last generations of a crowded
Earth. Ladies and gentlemen, don't let this new world order, this little lie, become something
that you pass on to your children, make sure that you tell them the truth about the sacred biology
of the world, the reverence that we should have for it. Do not let other men with machines control
your kids and stop all transfections in humans, because they are trying to eliminate the controls
with many needs necessary. I'm sorry for a little bit of disorganization today, not a lot of slides,
just a couple study halls today. I cut it in the middle, hopefully so that peer tube can handle
it a little better. But I think it's probably going to fail on the second one. We'll see.
And then we'll have tested the system a little bit more. Thank you very much for joining me.
You will see me again tomorrow. Intramuscular injection of any combination of substances
with the intent of augmenting the immune system is dumb transfection and healthy humans
is criminally negligent and RNA cannot pandemic.
And I'm wearing my Brooklyn number 11, which is actually Kyrie Irving. Kyrie Irving one,
he was on the Brooklyn. When he was on the nets, did not take the transfection,
he kept himself part of the control group. So although I do think that Kyrie Irving is a jersey
worth having, I actually think this is the Kyrie Irving jersey worth having, because for me this
is when he was standing up for the biology. And so shout out to Kyrie and I will see you guys again
very, very soon.