"Dysregulated Transmembrane Ion Gradients Highlight Cancer Invasion and Proliferation" and "Over the River and Through the Woods and into the Brain we Go: lupus autoantibodies offer a new pathway to treating brain tumors"

February 24, 2022

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Presentations by: Fahmeed Hyder, PhD and James Hansen, MD, MS

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00:00We have a number of people on
00:02the zoom already and I'm sure
00:04there will be more that the join.
00:07I'm very pleased today to have two speakers.
00:13Doctors, Hyder and Hanson.
00:15They will they will go in that
00:19order and so first is Pharmd Hyder,
00:22who is a professor of radiology
00:24and biomedical imaging and
00:26biomedical engineering.
00:28He received his PhD in
00:30biophysical chemistry from Yale,
00:32where he was also an associate
00:35research scientist and postdoctoral
00:36associate with Douglas Rothman.
00:38He's been in the faculty since 1999,
00:41seems hard to believe.
00:43You look very young and currently
00:46holds dual professor appointments
00:49in diagnostic radiology
00:51and biomedical engineering.
00:53Doctor Hatter is the director of
00:55the High field horizontal smallbore
00:57systems at Yale's MRI Research Center.
01:00He uses MRI.
01:01He uses magnetic resonance methods
01:03to map Physiology and chemistry
01:05that underlie brain function
01:07for early disease detection,
01:09but also for targeted drug
01:11delivery and monetary treatments.
01:12It's also the founder and director
01:15of Yale's quantitative neuroscience
01:17with magnetic resonance course enter
01:19the only NIH supported programmatic
01:21effort at Yale on neuroimaging.
01:23With Mr technologies.
01:24His work has produced over 100
01:27peer reviewed publications and
01:29he's written and edited books
01:32on functional brain imaging.
01:34I have to say something that I'm
01:37becoming increasingly interested
01:38as I age and he has received early
01:41career awards from various scientific
01:43societies and funding scientific
01:46agencies for me to pleasure.
01:48To have you.
01:49Thanks for being here and we
01:50look forward to hearing from
01:51you. Thank you, thank you very much
01:54for that generous introduction.
01:56I hope I can live up to that introduction.
02:00It's a pleasure to be here.
02:02So hopefully you can see the slide.
02:05Yep, perfect.
02:09So my topic today is about disregulated
02:12transmembrane ion gradients.
02:14I like cancer invasion of liberation.
02:17I believe the latter 4 words are probably
02:20more akin to a lot of the audience members,
02:22but I'm going to make the connection or
02:25try to at least before the the initial
02:28part of this title, which is about.
02:32So a little bit of.
02:34Preface I guess I'm interested
02:36in bringing the tablets in why?
02:39Because they tell us about the path weights.
02:42Which you know end field cells use
02:45nutrients to fill their function,
02:47but also growth.
02:49I'm interested in how cells work.
02:52What their made nutrients
02:54are in different situations,
02:55different states how they
02:57get these nutrients.
02:58That's actually a very important
03:00indicator of disease onset.
03:02A lot of fences,
03:04but also how they use these
03:06nutrients to generate energy,
03:07which is crucial for the
03:08brain but also to regrow.
03:11And disease and damage and so on.
03:14And all of these processes happen
03:17naturally for normal situations,
03:18but they also begin to malfunction.
03:22So these questions that I pose for myself,
03:27as well as my group, my movies and vibrators.
03:30These questions are fundamental for
03:33functional imaging of the brain because
03:35the energy demand for normal brain
03:37work is extremely high and that still
03:40is a very unique property of how.
03:43This organ varies from a lot of
03:46other buildings in the body,
03:47but these questions are also relevant
03:49in cancer and molecular energy
03:51of brain disorders in general,
03:53but especially concert,
03:55because this fundamentally is
03:56a disease of both,
03:58and both requires lots of fuel.
04:01Whenever you need to regulate
04:03to compete for fuel that is a
04:06medical question that it's tough.
04:08So that's my preference,
04:09but my main objective today is to
04:12talk to you about the importance
04:15of transparent votes.
04:16If they're crucially linked
04:18to cellular metabolism,
04:20and this property differs substantially
04:23between normal cells and cancer cells.
04:27Abnormal transmembrane protein and sodium.
04:30Specifically,
04:30I'll talk about protein and sodium gradient.
04:34Are there consequences of health
04:37physiological alterations
04:39occurring with cellular level?
04:41And the two primary things I'll
04:43focus on is the city and salinity
04:46of the interstitial fluid.
04:48These two quantities,
04:50reflected as a proton and sodium ions,
04:54respectively.
04:54They actually regulate central cellular
04:57functions in health as well as improved,
05:00especially cancer under
05:01their bark cancer tomorrow.
05:03Basicness,
05:04which is one of the hallmarks
05:06and even resistance to therapy.
05:08This is actually enhanced
05:10by acidic interstitial.
05:14Which is actually a consequence
05:16of popularity by policies.
05:18I will talk about this in a few minutes.
05:21But there's also recent
05:23discoveries from our work,
05:24and I work with and related groups
05:28that show that enhance proliferation.
05:31Which is also a hallmark of cancer
05:35is impacted by interstitial so,
05:39so we both these city and the salinity
05:42of legislation that is crucial.
05:45A little bit of background,
05:46I think that's the background
05:48is probably redundant,
05:49but for the sake of being bad
05:51protein and sodium lines are
05:53vital for numerous processes for
05:55maintaining blood pressure to fire.
05:58Only statically we maintain
06:00a very large sodium gradient.
06:03Almost two orders of magnitude,
06:05almost an order of magnitude.
06:06Sorry. Lending to a very strong
06:09trance member Radiant.
06:10Well, certainly, but similarly.
06:11For for time buildings,
06:13which is usually measured in terms of pH,
06:15which if you know it's
06:16just a log rhythmic scale.
06:18So even though the pH between interstitial
06:22and intracellular intracellular
06:24compartments are 7.4 and 7.2 in PS units,
06:29it's small.
06:30But in terms of actual concentration,
06:33it's again very large.
06:35There are various mechanisms with
06:37which are located the cell membrane,
06:39how regularly.
06:40A proton and certain levels to
06:43avoid these mishaps that are
06:45that sells try very hard before.
06:49First,
06:49Megan mechanisms that regulate
06:51transforming what time gradients
06:53are shown here in the parking form.
06:56Let me just go around the cell here first.
06:59Is this mechanism here carbonic
07:02anhydrase mine specifically?
07:04But these kinds of.
07:09Instruments are used to basically take out
07:12department dioxide and water generated by
07:16oxidation and take them out and it
07:19basically takes them out and comes
07:21to bicarbonate and protons and which
07:25signifies the intracellular space.
07:27Then there is these channels at BCS and NC.
07:30Is which abbreviations are shown here,
07:33but the second time I'll just
07:35go through the abbreviations.
07:36These bring in both bicarbonate
07:39and sodium and this one.
07:41Runs in bicarbonate itself, right islands.
07:44Both of these contributed to a contribute to.
07:48Changing of the intracellular field.
07:52Maximo carboxylate transporters
07:54are those kinds and city one and
07:58NC 3-4 working opposite days.
08:00They bring in assets for NCT one
08:04and take out assets for actually 4
08:08altering hydrogen ion concentration.
08:12It's all specifically.
08:15Important one entity,
08:17one sodium proton exchanger,
08:19takes like for every proton taken out,
08:22brings in every brings in a sodium and
08:25that can alter the full time as well.
08:30These are ATP and ATP dependent,
08:34vascular full-time ATP ace,
08:36where you're bringing in or taking
08:38out protons to a city public service
08:41place as well as these acid.
08:46Sensing line channels as well as these
08:50epithelial specific certain channels.
08:53They also are known to affect more
08:56time but as well as sodium because a
08:58lot of things different channels as
09:00sodium gradients and therefore beans
09:03won't be repeated because these are
09:05the channels that I just talked about.
09:07But there are other mechanism.
09:09This is a channel that I just
09:10showed in the previous slide.
09:11See energy as well as the
09:14sort of important change.
09:16Are all of these have a role in altering
09:19the sodium level in the exercise of space?
09:23This particular channel, the NSX,
09:25the sodium counseling exchanger,
09:27it's exchanges sodium for
09:29calcium between the compartments.
09:32It's interesting, it was recently discovered,
09:34but in cancer specifically,
09:36this is actually need to.
09:39In addition,
09:40there is this additional new channels
09:43mentioned here that are specific to
09:46sodium regulation across the department.
09:48The most important one of this is
09:50probably a well known as the support.
09:52The sodium proton switches exchanging
09:56of sodium and potent potassium at the
10:00cost of ATP to basically maintain a balance.
10:06To keep resting potential.
10:10The same so that the cell can
10:12actually keep on doing that.
10:14It's designed to do so.
10:15These are the mechanisms that are in play.
10:19Glucose metabolism is key to all
10:22of these processes and the healthy
10:24miracle as blood drains in both
10:27glucose and oxygen in the form of
10:30oxyhemoglobin glucose is its transport
10:33is expedited by these transporters,
10:35but oxygen goes through just
10:37passive diffusion into the cell.
10:41Just breakdown appointments through
10:42the bike clinic steps and then further
10:45breakdown in in in the prep cycle
10:48of generating large amounts of ATP.
10:51All of these processes,
10:52generally small amounts of protons
10:54but also carbon dioxide water which
10:57I just mentioned.
10:58Carbonic anhydrase has a has a role in in,
11:02in, in taking out these these these products.
11:05So this is a.
11:06Complete oxidation of glucose if you will,
11:09but even under normal conditions
11:12there is some.
11:16Less oxidation than expected from theory,
11:20where some of the glucose is actually
11:23shunted towards biosynthetic processes,
11:25but some of the invoices actually.
11:29Excluded in terms of lactate by the public.
11:33Feeds on CTV's that you
11:36have mentioned earlier.
11:37A lot of the protons generated are removed.
11:40As I said from these different
11:43mechanisms which I mentioned me
11:45cheese also about the heart races.
11:47But if cancer and the unhealthy durable the.
11:51A lot of dysfunction happens in
11:54terms of amount of bike policies,
11:56which is the amount of
11:59oxidation that happens.
12:00And therefore some of these
12:02there will be glycolytic.
12:04Steps are augmented to the point.
12:06Where these machinery's that I speak
12:09just mentioned with carbonic and
12:12hybrid sister upregulated in NCT 4?
12:15Is that up regulated to handle
12:17exclusion of lactate more efficiently,
12:21but also in terms of the NET,
12:25which is also up regulated to alter the
12:28whole time in the access center space.
12:31We vacuumed.
12:35Along with some of the same mechanism,
12:37the same machinery or additional machinery,
12:40you can also experience a lot of
12:43imbalances in a certain environment,
12:45but key player as I just mentioned
12:47is the mutation of the NSX,
12:49which happens which is most
12:51recently discovered in cancer,
12:53but also the fact that there
12:56is actually downregulation of
13:00oxidation within a cancer cells.
13:04Reduce potassium sodium is also an important
13:11role of causing these sodium imbalances as.
13:15And it's culture.
13:17So this is the unhealthy miracle and our.
13:21Our role is to understand and observe
13:24these the impact of the altered proton
13:27as well as transgender, Ingrid.
13:29So tumors are highly like little
13:31even when off, well oxygenated,
13:34and this aerobic glycolysis phenotype.
13:38Enables enabled by metabolism.
13:40Different nutrients generates
13:41lots of their civic products
13:44that are extremely deficient.
13:46For example,
13:46even carbon dioxide and water generated
13:49from oxidative metabolism are,
13:50you know,
13:51they contribute to certification
13:53of the interstitial space by these
13:55these anionic exchangers like.
14:00Invite.
14:03Interstitial acidosis actually
14:04helps with tumor cell invasion.
14:07They they actually did break
14:09the interstitial matrix,
14:11and they also promote angiogenesis
14:14while suppressing entry. Sense.
14:18So this is an example from Bob
14:22Giles work where it is shown that
14:26whenever there is use acidification,
14:28this is done in a culture system
14:32where there's reduce acidification,
14:35there is enhanced invasion of
14:38cells in terms of timber growth.
14:41This is an example where
14:42you study shown in situ,
14:44but in people evidence for this
14:46from our lab and other labs,
14:47so security is absolute.
14:51Impact with the altered transplant in
14:53sodium gradient normal cells maintain.
14:55Hyperpolarized membrane potential.
14:58But before I see member interventional
15:01and non excitable cells is linked to
15:06their proliferation of point to this in
15:10in the next slide a little bit more.
15:12In fact, even in mean cells consensus
15:15in the interstitial in the loop.
15:17Therefore,
15:18salinity of the interstitial and
15:20intracellular compartments may be crucial.
15:23As has been pointed out by recent.
15:26Work that may be important for
15:29early diagnosis of cancer,
15:30but also maybe contracting
15:32in cancer treatments.
15:34So this point, this light points this out.
15:36This is the scale of membrane potential.
15:38On the bottom are these
15:41non proliferating cells?
15:42These are real cells,
15:44neuronal cells.
15:44They have a very hyperpolarized
15:48membrane potential equation which
15:50takes into account sodium and
15:52the poor at concentrations up.
15:54Here are the perfect.
15:55Or looks writing cells which are
15:57non tumor cells but the membrane
15:59potential measure of tumor cells
16:01in the rieti of different kinds.
16:03You see they right up here in the
16:07depolarized level. So in essence.
16:10What a lot about solutions from
16:13various groups have shown is that.
16:18The week transmembrane sodium
16:20gradient maintaining a depolarized
16:22membrane potential is actually
16:24necessary for these cells to survive
16:26in their altered environment that
16:28they create for their survival.
16:31Update at the.
16:32Cost of normal cells that they're replacing.
16:36Whereas you know these normal cells,
16:39especially in the brain,
16:40maintain strong,
16:41transmembrane certain buildings
16:42and they may do so by maintaining
16:45their hyperpolarized.
16:49So these are the hallmarks of cancer,
16:52our goal, and my goal in the next few
16:55minutes is to show that you can employ
16:57sodium and proton imaging methods
17:00to actually observe 2 properties,
17:04invasion and proliferation of cancer cells
17:07by using proton and imaging perspective.
17:11We talked a little bit about that.
17:13I'm sure this method is quite
17:15familiar with most.
17:21Slash house light.
17:38Turns out that.
17:45In like one step. Slide disappeared, yeah.
17:49I realized that it just didn't like 1 slide.
17:55Do you wanna stop sharing and start
17:56again or is that what you're doing?
17:58Yeah, I'm trying to do that right now, OK?
18:01OK, so I'll skip that slide.
18:10And see if I can get from that.
18:15So the slide that I unfortunately had that.
18:21You know it bounced me out of PowerPoint.
18:24Basically this describes a key late.
18:27That is the design structure of most contrast
18:30agents that is used in the preferable
18:32design is where you take a gadolinium.
18:35Mine is very car magnetic and you
18:38use that to reduce its toxicity
18:42Yuzuki late with donating.
18:44Components like oxygen atoms that would
18:47provide the stability for conjugation,
18:50reducing discovery toxicity
18:52of the metal iron.
18:54But discriminating iron is some of the
18:57key of the types of imaging observations
18:59that I'm going to show you about.
19:02So this is essentially a proton
19:04spectrum of a key late like this.
19:06This is a it's called appear TP molecule.
19:09It's basically a molecule that
19:12contains multiple phosphinate groups,
19:14much like phosphonates that exist
19:16in molecules like ATP and ADP.
19:18So this is a very
19:20common diamagnetic range of signals that
19:22you would see emanating for different.
19:25Protons, the non exchangeable protons that
19:28exist in these carbons as well as the
19:31carbons existing in these pendant arms.
19:36By the way, I point out that these
19:37phosphates on this on a molecule like this.
19:40These protons exchange with protons
19:42of water and that kind of exchange
19:45is actually a a a pH mediated
19:48and and and therefore they are.
19:52These molecules are essentially
19:54create sensor much like ATP and
19:58ADP is using phosphorus and.
20:00So if you now collate this,
20:03take this molecule and and complexes
20:05with the paramagnetic metal.
20:07In this case,
20:07I'm not using gadolinium,
20:08but a fully am I and you essentially
20:12caused this large expansion of the
20:14same chemical shift that you see here
20:16within a few partner million and that's
20:19expanded by hundreds across people.
20:21So these signals have no potential
20:24overlap with other types of signals
20:27that you may observe in vivo.
20:29And this is the water signal.
20:31And so this hyperfine shifted
20:33signal has very unusual properties.
20:36Basically, all it means is that
20:38you can observe them quickly.
20:39You can observe them under very
20:43precarious in vivo situations that we
20:46may experience the relaxation times are
20:49so short because of the paramagnetic
20:51environment that that these photons
20:53are in create a very sensitive scenario.
20:57That's most, importantly,
20:58that the chemical shift of these signals,
21:00not the amplitude,
21:01of these signals.
21:02But how these signals are
21:05located and how they shift?
21:08Is the readout that we observe again,
21:10so this is a molecule which is the sensor.
21:13So if you change the
21:15environment of the sensor.
21:17The structure of the molecule will change
21:19and as the structure of the molecule changes.
21:22What you then read out that structural
21:25changes from the chemical change.
21:28And so this is encapsulated here.
21:30This is a molecule that we
21:32are using as our sensor.
21:33These are the types of signals that we
21:37observe using this as an example 86 proton.
21:40It has a pH sensitivity,
21:42but obviously any molecule reports
21:43environment by changes like these
21:45molecules have or molecules like this
21:47have very very short relaxation times.
21:49So you can observe them quickly
21:51and therefore imaging these have
21:53been possible and the readout from
21:56them in terms of chemical shift is.
21:58Not confounded by other overlapping signals.
22:02So this is an in vitro example of this
22:05kind of work from my colleague Dan
22:08Employment and then this method is.
22:11Paul by sensor imaging,
22:13redundant deviation shifts, birds for short.
22:15Essentially showing you that in these
22:18two phantoms which have very different pH,
22:21you can read out the pH with high confidence
22:23even if you change the temperature.
22:25So you can essentially
22:27give these two things out.
22:29Simultaneously,
22:30the key thing is is that this is
22:33not like most imaging contrast.
22:36This contrast is not based on
22:39the amount of agent that you have
22:42in in the given compartment,
22:44but really their chemical shift.
22:45So it's it's a readout that's
22:47independent of concentration,
22:48and so using this method we've
22:50been able to apply it to look at
22:53the cancer environment,
22:54specifically the the pH in the extracellular
22:57space, and a variety of different.
22:59Scenarios these these are examples
23:02from breaking specifically shown for
23:05two different types of tumors here,
23:06but you can note and appreciate the acidity.
23:09Acidity within the tumor poor as shown here.
23:13Core being identified by the line
23:18that shows up in the MRI contrast.
23:21But you can also appreciate this
23:23extended area of a certification
23:24for this tumor and not that tumor,
23:27and that has to do with the
23:29how evasive this tumor is,
23:31and using cellular markers,
23:33you can corroborate that funding,
23:35so this goes back to the
23:37point that our pH readout,
23:39so this is a case of a a
23:41very non invasive tumor,
23:42versus this one is tumors invade and this.
23:47Invasion if you will.
23:49I we believe is essentially preconditioning
23:53for the expansion of this tumor within time.
23:56For these two specific cases,
23:58here in the 2nd and R2, but not the help.
24:03So talked about a little bit about what
24:06we do with the Proton imaging part to
24:08read up pH and that uses the proton nucleus,
24:12right?
24:12This is a very highly sensitive
24:15nucleus to observe because with the
24:17hygiene gentlemen medical issue.
24:19But what I'm going to talk about next
24:21is the sodium imaging part which has
24:25a lower gyromagnetic ratio but aided
24:27by the fact that we are looking at.
24:30You know our product nucleus we are.
24:33So we can benefit a little bit from the
24:36certain relaxation times of this nucleus.
24:39So that's kind of where we are.
24:42Our basic rationale for this type of
24:45experiment is that the problem that
24:47we face is that the sodium signal,
24:49whether looking at the the interstitial
24:53or the intercellular pool,
24:55signals they overlap.
24:57They're not chemically different,
24:59and therefore the total signal
25:00you observe is is a representative
25:03of these two types, etc.
25:05Our expectation is that somehow we can
25:08separate the two and that separation.
25:11Will allow us to look at things
25:13like the transmembrane great.
25:16So the idea stemmed from work
25:17that existed in the field,
25:19but the advanced it in the last few
25:22years is that if we have a polyanionic.
25:26Our magnetic agent,
25:27much like pet agents that I've talked about.
25:30And we take a sodium ion because
25:33of its negative charge.
25:34It's attracted to it and there
25:37is superbound bounding of some of
25:41these ions because of the negative.
25:44Target of this agents and we have
25:46some exchange of free sodium with
25:49this sort of bound sodium,
25:51and if that exchange happens.
25:55Fast enough that we can reflect the
25:58shifting of the sodium signal from
26:01a from this process to reflect the
26:04true 2 compartments of service.
26:06So in essence,
26:07what we do and this is our rationale take.
26:11Before we add an agent like this,
26:13this is the total signal we will look here.
26:16And when we inject or introduce
26:19an agent like this because it's so
26:22negative and it can attract positive
26:24recharge sodium ions towards it,
26:26we can separate these two sodium
26:30compartments.
26:31And this is a theory which we've
26:33contributed to, and a key.
26:34Two key factors that we point out in in
26:38this theoretical and practical demonstration
26:40of pages like this is the fact that.
26:43There is a certain bound fraction of sodium
26:46towards these agents that's important,
26:48but also their exchange and these two
26:51mechanisms have NKX actually contribute to
26:54how much the sodium signal will be shifted.
26:58So the shift ability factor,
27:00but also how these signals could be broadened
27:03because of their kind of like nature.
27:06So, uh, another key thing that we point
27:08out is that these shifted signals
27:11brought in signals of of of sodium
27:13that we will observe is dependent
27:16on the sodium concentration on the
27:19on the agent concentration.
27:20And here we are limited by the fact
27:23that we need sufficient amount of
27:25this agent present to process certain
27:28for the broadening and therefore
27:30this is something that will act as
27:33a benefit for certain sensor.
27:36But it was not necessary for
27:38them for the poor concepts.
27:40So our expected idea for these
27:42experimentation is that before we add
27:45the agent, we're seeing one single,
27:46and after we add this agent,
27:48we're gonna separate the signal.
27:50Now you see three and I point out
27:53three because you also have sodium.
27:55Plenty of sodium also in the
27:57blood department.
27:58But because the the blood department is much
28:01smaller than the interstitial compartment,
28:03these representative amplitudes
28:05of these beats are depicted.
28:09So in vivo inside a tumor voxel.
28:12This is what we see.
28:13This is before the agent.
28:15It's been separated here.
28:17And outside of the tumor,
28:19we see something that's similar but
28:21to the shifting and the broadening
28:23is to some lesser extent.
28:25So this is kind of what we
28:27typically do nowadays,
28:28and this is work by Mohammad Khan who
28:30did his thesis work on this project.
28:32Is is this is the conventional
28:34location of the tumor.
28:35We inject the agent to
28:37identify the blood compartment,
28:38interstitial compartment,
28:40interstitial intracellular compartment
28:42in terms of sodium.
28:44We integrate these signals to
28:46get maps like this,
28:47and using a combination of these two,
28:49we can get what's called
28:51an endothelial gradient.
28:52This is asserting gradient representing
28:54the and the feeling department.
28:56And this is the critical one
28:57that we will talk about,
28:59which is the transmembrane gradient
29:01which is obtained from the intracellular
29:04and the interstitial signals also.
29:06And we we can now map this into ID
29:09to get the transmembrane gradient
29:11how it appears within the tumor.
29:14I point out there's these blobs of high
29:17intense signal represents the ventricles.
29:20Ventricles hasn't has a
29:22lot of sodium in them,
29:24and that has variety of them applications.
29:28Or other disease disorders as well,
29:31but the key thing is that we can
29:33obtain a clear readout of the
29:36transmembrane gradient shown for two
29:38different three different tumor types,
29:40and so this is a pretty uniform and
29:43vigorous observation that we can now make.
29:46We even began to employ these
29:48methods for variety of
29:50different treatments.
29:53And and so this is a very unique entry.
29:57Genic antiangiogenic treatment and
29:59what is shown here is a comparison
30:01of what's your afternoon, does it?
30:04It it blunts it.
30:06It sort of impedes that's in the growth
30:08which is most significant within two weeks.
30:11And this kind of effect can be
30:13read out by the teenage as well
30:16as the transmembrane gradient.
30:17As you can see the sorafenib against
30:20renormalize the pH with treatment and.
30:24As well as normal, you know.
30:28Strengthening the sodium transmembrane
30:30gradient upon treatment even
30:32the Chamber making it similar
30:34to the normal tissue as well,
30:36so I hope that's been able to show
30:39within the last few minutes is
30:41that we now have a quote which can
30:44sense both quote photon and sodium.
30:46A lot of credit to both.
30:49I'm sharing design quotes of
30:52this particular quote,
30:54and quotes like this,
30:55which with proton imaging can be a very
30:58powerful proton sensor for pH imaging,
31:01but also a sodium sensor in terms
31:04of its its signal shifted.
31:07It it can be also used simultaneously.
31:11So I hope I've what I've
31:12shown is that all cells,
31:13not just excitable neurons and muscle,
31:16generate and receive by electrical
31:19signals that are encoded.
31:21Within changes in the transmembrane
31:24potential and the iron fluxes that
31:26occur at the cell membrane and
31:28these things happen regularly,
31:30you know on timescales of milliseconds,
31:32seconds to even days.
31:34But these are inextricably
31:37regulated by catabolism,
31:39and that is the connection that it is is.
31:44This is needed to find the proper readouts
31:48of various treatments that we see.
31:51And I, I think this is another way of
31:54saying this is that we need advancing
31:56before imaging methods to assess the
31:58non genetic by physical aspects of
32:00tumor microenvironment that regulates
32:02balance between normal growth
32:04but also the disorganization that
32:06happened with most solid solid cancer.
32:11But importantly,
32:12simultaneous imaging invasion and
32:15qualification can hold a promise for
32:18early cancer diagnosis and tracking
32:21therapies. From chemotherapy to two.
32:25There's always grateful for
32:27tonight support and this work is
32:29unique collaboration among many
32:31colleagues within our group.
32:35Khan and John Walsh,
32:38PhD and MD,
32:39PhD students are involved in a
32:40lot of this work,
32:41but also by colleagues in
32:43terms of post starts.
32:44You're still present Doctor
32:46Kumar Mishra and and the high
32:48leverage but also my colleagues.
32:52Within radiology and
32:54surgery that department so.
32:57And I thank you for your.
33:01Your attention.
33:03Thanks, thanks very much it.
33:05I think that was great.
33:06You mentioned chemotherapy and
33:09immunotherapy, and you know,
33:10our next talk is from a radiation
33:12oncologist and one wonders whether this
33:15might also help identify tumors and
33:17tract tumors that are being irradiated.
33:20Since, as he well knows,
33:23it is that his doctor Hanson.
33:26It is sometimes confusing
33:27after someone's been treated,
33:28whether there's active tumor or or not.
33:31So without further ado.
33:33Guy James Hansen is an associate
33:36professor of therapeutic radiology
33:38and chief of the Gamma Knife
33:40program in therapeutic radiology.
33:42He received his medical degree
33:44from UCLA School of Medicine and
33:47Masters degree in biochemistry and
33:49Molecular Biology at UCLA as well.
33:51Doctor Hanson,
33:52Clinical area of expertise is gamma
33:54knife stereotactic radiosurgery for the
33:56premium of CNS tumors had neck tumors,
33:58lung cancer as well as lymphoma,
34:00skin,
34:01GI and GUI will bet that he spends more
34:03time on CNS disease than other things.
34:06But he can tell us his research focus
34:08is studying the interplay between
34:10autoimmunity and malignancy and
34:12attempting to harness and optimize select
34:14autoantibodies to use against cancer.
34:16James, thanks for.
34:18Joining us.
34:19Thank you and thank you
34:21for that introduction.
34:21And yes, if if you can explain
34:23my clinical career to me,
34:25that would be great.
34:26I would love to. But it's hot.
34:30Alright, so let's see,
34:31can you see my slides and hear me?
34:34Yeah, it's in that presentation
34:35now that's great. Yeah
34:36alright. Here we go.
34:38So I'm gonna be talking today about
34:40my particular interest in using
34:43lupus antibodies against cancer,
34:45and in this case, how we can maybe use lupus
34:48antibodies against brain tumors and so.
34:51Already, I probably have lost half
34:53of youth that are just saying,
34:54well, that's impossible.
34:56Antibodies can't do that.
34:57But just give me 20 minutes or so,
34:59give me a chance.
35:00I I think that there's something
35:01here that's worth talking about.
35:03Do you have some disclosures?
35:04I'm a consultant.
35:05I have grants from inventor
35:07on a patent licensed by this
35:09company called Patrys Limited,
35:11who has licensed the DEOXY mab technology.
35:14So with that said, let's jump head first in.
35:17I don't think there's any
35:19secret that antibodies have
35:21revolutionized our approaches.
35:22In modern day two treatment of cancer.
35:25But I think it's important to recognize
35:27that all the antibodies that we really
35:29rely on in the clinic right now are
35:31targeted towards extracellular targets.
35:33So things like surface receptors
35:36or circulating growth factors.
35:38But doggone it seems like there
35:40are so many intracellular antigens
35:41that if we could just get an
35:43antibody in there to engage,
35:45we could add an entire new
35:47dimension to immunotherapy.
35:48But the dogma has always
35:50been that that's impossible.
35:52Antibodies don't penetrate live cells.
35:56Now this is where critics will often
35:57interrupt me and say that's not true.
35:59We have antibodies that are already in
36:01clinical use that are internalized.
36:03What about cats?
36:04I love the TDM ones and I I would
36:07say I say DNA.
36:08That's not what I'm talking about.
36:10I'm talking about an antibody that can get
36:12in and engage its actual native antigen.
36:15In these ADC constructs,
36:17these antibodies are still looking
36:18for surface receptors like her
36:20two in this example and then the
36:22antibody gets eaten up and destroyed
36:24in the endosome and lysosome which
36:26is great for this mechanism because
36:28that's what we want,
36:28we want that drug to be released,
36:30but that doesn't utilize the exquisite
36:33binding specificity of an antibody
36:35against intracellular targets.
36:36So I think we need an antibody
36:38that can get into cells and not get
36:40stuck in those Endo celebs, but.
36:42How are we going to do that well?
36:45I guess we could try to invent one.
36:46We could stick things like cell
36:49penetrating peptides onto antibodies
36:50like that at peptides and such,
36:52and that's been tried,
36:54but those tend to still get stuck
36:56in the endosomes.
36:57I guess we could try gene therapy, but boy,
36:59that's got all kinds of challenges.
37:01I'm going to leave that to other people.
37:04Maybe if we can just find a naturally
37:06occurring antibody that penetrates
37:07cells and use that as a platform to
37:09teach us how to invent these antibodies,
37:11that seems the most appealing to me.
37:14But anybody have any idea where we're
37:17going to find an antibody like that well?
37:20How about in lupus?
37:22Systemic lupus erythematous?
37:24It is the prototype autoimmune disease.
37:27Patients suffer widespread tissue
37:29destruction and inflammation
37:30as their immune
37:32systems recognize their
37:33own cells and tissues.
37:35And one of the laboratory hallmarks of
37:37lupus is the presence of circulating
37:39autoantibodies that are reactive
37:41against the patient's own DNA.
37:44Now those antibodies are a big mystery.
37:46We still don't know exactly how they
37:48contribute to lupus pathophysiology,
37:50but remarkably it is now finally
37:53accepted that a small percentage
37:55of them actually have the ability
37:57to cross through membranes and
37:59penetrate into live cell nuclei.
38:02Well, hey, so there we have it right.
38:03We have a source of naturally
38:05occurring cell penetrating antibodies.
38:06We can use those for all kinds of therapies,
38:08right?
38:08Well, hold on now everybody
38:11we're talking about Lupus right?
38:13At last I checked Lupus is still
38:15in the textbooks as a disease.
38:17And indeed,
38:18a lot of these cell penetrating lupus
38:20autoantibodies are just broadly cytotoxic.
38:22And there there wouldn't be any
38:24benefit to giving them to a patient.
38:26But thankfully,
38:26that's not true for all of them.
38:28There's an antibody called 310,
38:31which is the hero of this story.
38:33It was discovered in the early
38:341990s at UCLA by Richard Weisbart
38:36who's pictured there at the left,
38:38along with his technician Grace Chan and
38:40his great colleague Robert Nishimura.
38:42Their great,
38:43great friends and colleagues.
38:45What makes 310 so remarkable is that
38:48it was isolated from a lupus mouse.
38:50It penetrates extremely effectively
38:52specifically into the nucleus of live cells,
38:55and so it does not go through endosomes,
38:57and it does not kill or is not
38:59toxic in any way to normal cells.
39:02So now we've got our chance.
39:04Now we have an opportunity to use a
39:07platform antibody that penetrates cells.
39:09And in fact,
39:10as we dive a little deeper into
39:13how this antibody really works,
39:15it turns out this antibody is really
39:17well situated for targeting things
39:19like tumors or sites of tissue damage.
39:22And that is because part of its
39:25mechanism of penetration is dependent
39:27on presence of extracellular
39:28DNA or nucleosides in the area.
39:31So what we're showing on the left
39:32here is something we call the
39:34three E 10 bullseye effect of the.
39:35The dark stain represents where
39:37the antibody is, and as you see,
39:39that dark stain is getting lighter
39:40and lighter as you get further
39:42out from the center,
39:43and that's because there's a
39:44dead cell there in the middle.
39:46And it's releasing DNA into its
39:47surroundings and helping the antibody
39:49penetrate the live cells that are
39:51closest to it, but less effective.
39:52As we get further out.
39:54And in the middle we showed we can
39:56reproduce this in the laboratory
39:57just by adding DNA to the antibody.
39:59At the bottom we allow the antibody to
40:01penetrate 100% of the cells in the culture,
40:03so the DNA has to be there.
40:06And then if you take this antibody
40:08to mice and you give it to mice that
40:10don't have any tumors or damage,
40:12you don't see the antibody really
40:14going anywhere.
40:15But if you give it to a mouse with a
40:17tumor that is necrotic and is releasing DNA,
40:20you do see the antibody
40:21localising to that tumor.
40:22That's what's shown in the top right.
40:24The brown stain are the nuclei
40:26penetrated by this antibody.
40:28So what on Earth is going on here?
40:30Why is this antibody using
40:32DNA to penetrate cells?
40:33How is it doing that?
40:34Well, that's a little bit of a longer
40:36story than we have time for today,
40:37but just to jump to the punch line,
40:40it is using a specific nucleoside
40:42salvage pathway to get into live
40:45cells that are salvaging DNA and
40:47nucleosides from their surroundings,
40:48and that transporter is called ENT two.
40:51It is my best friend transporter
40:53in the whole world stands for
40:56equilibrated Nucleoside transporter 2.
40:59So in order for 3:10 to cross membranes
41:01to penetrate into cells and nuclei,
41:03you have to have two things.
41:05The cell has to express ENT two and
41:08there has to be DNA or nucleosides
41:10around that cell so that the antibody
41:13can bind to the nucleosides and then
41:15follow them through ENT 2 into the cell.
41:18And that's why the antibody likes
41:20to preferentially accumulate
41:21in vivo insights of damage,
41:23like tumors where DNA is being released
41:26and ENT two is salvaging salvaging
41:28the DNA and along with it the 3:10.
41:31Networks for injury sites as well,
41:33and that's what we've seen
41:35in multiple studies.
41:37For example,
41:37moving from left to right,
41:39if you take the three E 10 antibody and
41:42you link it to a heat shock protein.
41:45And then give it to mice or rats.
41:47Actually that have had strokes.
41:48You find the antibody gets the heat
41:50shock protein into the ischemic brain and
41:53improves neurologic function and recovery.
41:55In the middle here we're showing
41:58heart attacks in rabbits.
41:59The 3:10 antibody finds the site
42:01of the heart attack and delivers
42:03the heat shock protein.
42:04It works remarkably well.
42:05And on the right we're looking at tumors.
42:08You take the antibody and you fuse it to P53.
42:10That protein from long time ago
42:12and it absolutely localizes the
42:13tumors and shuts them down.
42:15So 310 has great potential as a delivery
42:18vehicle for tumors or sites of damage,
42:21and there's now more companies
42:24looking at this than I can even count.
42:27But that's not all the antibody does.
42:29It turns out that it does more.
42:31It's not just a delivery agent.
42:34When the antibody penetrates into the
42:36nucleus of a cell, it will bind DNA.
42:38But if it has its choice,
42:40it will preferentially bind
42:42a DNA that's broken.
42:43And then it's going to mess around with
42:45DNA repair so it blocks base excision,
42:46repair,
42:47and rad 51 mediated molega's recombination.
42:50And that means it can make cancer
42:52cells more sensitive to DNA
42:54damaging therapy like certain
42:56chemotherapies and radiation.
42:57But even more significantly,
42:59if the cancer cell or the tumor has a
43:03pre-existing defect in DNA repair due
43:05to a mutation such as Bracco or P-10 loss.
43:08The antibody doesn't need radiation.
43:10It doesn't need chemotherapy.
43:11It will kill that cancer cell
43:13by itself by causing persistence
43:15of DNA double strand breaks,
43:17but that doesn't happen in normal
43:19cells that have intact DNA repair,
43:21so we have a selective toxicity.
43:244HR deficient tumor cells with this antibody.
43:28So we reported that quite a while ago,
43:32and then we asked the question,
43:33is it just 310?
43:34Is this just something magic about
43:363:10 or is this true for other
43:37cell penetrating antibodies?
43:39And it turns out indeed
43:40it's not just three ten.
43:42There are antibodies that
43:44penetrate cells and cut DNA and
43:46kill the HR deficient cells.
43:47So there's a pattern emerging here
43:50where lupus antibodies some lupus
43:52anti DNA antibodies seem to be
43:53toxic to HR deficient cancer cells,
43:56and so then that led us to start
43:58asking some questions about.
44:00What does this mean for lupus
44:02and and cancer risk in lupus?
44:03And I'm not a rheumatologist.
44:06I did not know,
44:07but we started reading and looking.
44:08And overall,
44:10cancer risk is increased in lupus,
44:13but it's driven mostly by
44:15haematological legacy.
44:16So if you back up and you say, well,
44:18let's go tumor type by tumor type,
44:20you get a surprising finding in
44:23that breast cancer occurs at a
44:25lower than expected rate in lupus.
44:27And this has been widely recognized
44:28for years,
44:29but no one can figure out exactly why.
44:30There's no clear associating factor.
44:34But if you look a little deeper,
44:35it looks like it's the triple
44:37negative breast cancer that is
44:38specifically suppressed in lupus.
44:40And we do know that triple negative
44:42breast cancer is associated with the
44:44braknis phenotypes and the HR deficiency.
44:47So then we start to think,
44:48well, if that's true, like why?
44:50Why aren't lupus patients making these
44:52triple negative breast cancers as much?
44:54Well,
44:54we know about these DNA damaging
44:56autoantibodies that are killing
44:57these HR deficient cells.
44:58Is it possible that lupus anti
45:00DNA antibodies actually are
45:01protective against breast cancer?
45:03And if so,
45:04maybe we can re engineer them
45:05and use them to treat triple
45:07negative breast cancer and so?
45:09We're excited about this.
45:10We sent it in as an opinion letter.
45:12I thought I was extremely clever.
45:14I was very impressed with
45:15myself when I called it.
45:17The lupus butterfly effect,
45:18because lupus is the symbol of that.
45:21Sorry, the butterfly is a symbol of lupus.
45:24The butterfly effect is a symbol of chaos.
45:26There's chaos and immunology.
45:28Chaos of using lupus
45:30antibodies against cancer.
45:31I thought it was great title and then
45:32rear said now you haven't proven it.
45:34You gotta call the theory so it was
45:36published as the lupus butterfly theory,
45:38but I will forever want to
45:39remember it as the lupus butterfly
45:40effect theory hypothesis.
45:41Postulate whatever you want to call it.
45:45But we do need to prove it,
45:46so we published that in 2016
45:48and we started asking around.
45:50Anybody can help us with
45:52epidemiology 'cause we have no idea
45:53what we're doing in that regard.
45:55And it turns out Lupus is
45:57rare enough that it's a.
45:58It's a challenge to actually prove
46:00the association between lupus
46:02antibodies and breast cancer risk.
46:04But not an insurmountable one because
46:06John Hopkins was able to do it and
46:08they published it just last year.
46:09So this. Kind of blew my mind
46:11and I was very exciting to see
46:13this the Hopkins Lupus cohort.
46:15They were able to treat.
46:17Sorry they were able to analyze
46:192000 plus lupus patients that
46:21entered their cohort without a
46:23cancer diagnosis and then evaluate
46:24their risk of breast cancer.
46:28Over the years and they were able
46:30to associate that with the patients,
46:32laboratory studies and anti
46:34DNA antibody profiles.
46:35And we do see finally proof that
46:38there is an association between
46:41anti double stranded DNA antibody
46:43positivity and that reduction in
46:45breast cancer risk patients that make
46:47those anti D antibodies have 45%
46:49reduction in the breast cancer risk.
46:52And if you dig even deeper and you
46:54stratify the patients based on the amount
46:56of those antibodies they are making,
46:58the low producers did not have any
47:01reduction in breast cancer risk.
47:02But the high producers had
47:04a 59% reduction in risk,
47:06so I've I'm taking this as
47:08backing up my theory.
47:09I think the lupus butterfly theory can be
47:11called the lupus butterfly effect now,
47:12so I might ask for a revision
47:14to that article.
47:15Although it's been five years here.
47:17But regardless,
47:18it's it seems like we're learning
47:20something about lupus antibodies
47:21and at least breast cancer,
47:23and it gives me more confidence
47:25in what we're doing and trying
47:26to reengineer lupus antibodies
47:28to treat triple negative breast
47:30cancer and other tumors.
47:32So what are we doing in that regard?
47:33Well? Again.
47:35I remember and I recognize 310,
47:39although it's technically
47:40safe in normal cells.
47:41It's still a lupus anti DNA antibody.
47:44And you don't want to give
47:46anybody lupus like symptoms.
47:47So we gotta rethink this a little
47:49bit before we start taking
47:51this to clinical trials.
47:52The last thing you want is a lupus FC
47:54region being administered to patients,
47:56because the FC is going to activate
47:59compliment and ABC and all kinds of madness.
48:01But the good news is, 310 does not
48:03care whether or not it has the FC tail.
48:06It'll penetrate cells with or without the FC.
48:08It'll bind DNA and inhibit DNA
48:10repair with or without the FC.
48:12So first thing we've done, cut it off.
48:14There is no more FC tail,
48:16the danger is gone.
48:17And that's how we make what we call
48:19a single chain variable fragment,
48:21SCFE.
48:21It's only the variable sequences
48:23of the light and heavy chains.
48:26And that's been optimized to
48:27increase the affinity for DNA.
48:29And then we stuck a couple of those
48:31together to make a dye single chain
48:33fragments to bump up the avidity for DNA.
48:35And that works really well against HR
48:37deficient cancer cells and tumors.
48:38And that's the product that was
48:40finally licensed by biotech company
48:42Patriss as we talked about who
48:44then has helped with funds to
48:45allow us to humanize and demonize
48:47and further optimize the CDR's.
48:49To develop the antibody,
48:51I would like to now introduce
48:53named Deoxy Mab one or DX1 deoxy
48:55mab 'cause DNA mab for antibody.
48:58Yeah,
48:58you get it.
49:00DX1 is well if ENT two is my
49:03favorite transporter DX one is
49:05probably my favorite antibody,
49:06although there is some other ones that
49:09are kind of catching my attention too.
49:11It works great,
49:11penetrates so clearly into the
49:13nucleus we see triple negative
49:14breast cancer cells on the left and
49:16breast cancer brain Mets else on the right.
49:18It's killing the cells,
49:20it's sensitizing to radiation,
49:22but it's still leaving normal cells alone.
49:24Just what we want and we are
49:26excited to move this towards
49:27clinical trial testing.
49:28But wait a second did I?
49:30Am I saying something about
49:32brain Mets cells but?
49:36Am I arguing that we could maybe
49:37be using this to treat brain Mets?
49:38This is an antibody, right?
49:39I mean this, the story is
49:40already kind of far fetched, but.
49:42Brain Mets well, The thing is one of
49:44the key biologic markers that predict
49:46sensitivity to our antibody is loss
49:49of P-10 even more so than bracket for.
49:51And I know that all the DNA repair
49:53experts are probably jumping
49:54up and down right now saying,
49:55well the P-10 HR link is not
49:59completely proven and fine, but.
50:01Cells at a P-10 deficient are
50:04killed by this antibody and brain.
50:06Mints often exhibit P-10 loss even when
50:09the primary tumor is P 10 positive
50:11and that's either evolution towards
50:14metastasis or secretion of P-10
50:17suppressive micro RNAs by astrocytes.
50:20It's also worth noting P-10 is lost
50:21an awful lot in primary GBM, so.
50:23Maybe we've got treatment for GBM as well.
50:28But I hear what you're saying.
50:30Brain Mets brain tumors with an antibody.
50:34I I don't understand what the
50:35why the skepticism.
50:36I mean,
50:37all we need is DNA and nucleosides
50:39and the E NT 2 transporter right?
50:41It seems like the antibody should be able
50:43to get into the brain tumors just fine it.
50:45Oh oh right.
50:49The blood brain barrier.
50:53I guess that does pose something of an issue.
50:57Maybe I should have thought this through
50:59before I signed up for this talk.
51:00I guess it's too late to back out now though.
51:03If we look really closely at the
51:06blood brain barrier and we look in
51:08at the brain endothelial cells.
51:10Whoa, there's my buddy.
51:13E and T2. The luminal surface
51:15of the brain endothelial cell.
51:17And it actually regulates nucleoside flux
51:19into and out of the central nervous system.
51:22So wait a second.
51:23This is all starting to come full circle now.
51:26I think I'm arguing that our
51:28antibody DX1 can get over the river,
51:30the bloodstream, and through the woods.
51:33The blood brain barrier and
51:34into the brain tumor.
51:35We will go where the antibody will
51:38then bind DNA and inhibit DNA repair
51:41and kill those brain Mets and also.
51:44The possibly last owner.
51:48And we are seeing that this indeed is
51:51working out very well season. Sorry.
51:55We are finding that DX1 does cross transwell
51:58models of the blood brain barrier and it
52:00does so in an EMT 2 dependent manner.
52:02If you shut down EMT 2 the antibody
52:04can't get across in the middle panel.
52:06Here we're showing mice with
52:08orthotopic brain tumors.
52:09GBM in this case.
52:11And we gave the antibodies.
52:13We gave the mice antibodies labeled so
52:14that we can see it on imaging and the
52:17antibody gets into the brain tumor great.
52:19But not if we treat the mice
52:20with an inhibitor of ENT two,
52:22so you shut down ENT 2,
52:23the antibody can't get into the brain tumor,
52:25so it's working like we think.
52:27And it seems to actually matter.
52:29It makes a difference.
52:30The panel on the right.
52:31These are mice treated with the antibody
52:34with P-10 deficient patient derived GBM.
52:37DX1 by itself.
52:38No radiation.
52:39No chemotherapy significantly suppresses
52:41the tumor growth and extends survival.
52:49There we go. Brain Mets are a little
52:52harder to study, and GBM in mice.
52:54Just implant the tumor intracranially
52:57and watch and it goes, but that's not
52:59really fair for a brain met model.
53:01So the way we study brain Mets has been
53:03taught to us by our good friend and
53:06colleague Jangling Zhao and mastered by
53:08research associate my lab benedet caffari.
53:10These are not easy experiments.
53:14Breast cancer cells are injected
53:15into the hearts of the mice to allow
53:18them exposure to the circulation,
53:19and we use a brain seeking some type of
53:21the cancer cells to go to the brains.
53:24And you can then track those based
53:26on their signal on serial imaging.
53:28So if we give the mice brain mats and
53:30we treat them with tail vein injections
53:32with control, or the antibody,
53:34just the antibody, no radiation,
53:36no chemotherapy.
53:37I think that picture up in the
53:39top right says it all.
53:41It suppresses the brain
53:42Mets quite phenomenally,
53:44and it does also extend survival,
53:47so this is looking really, really good.
53:49But now I start to hear the words
53:51again in the back of my head.
53:52I don't know if anybody else can hear them.
53:54Do we really need a new
53:56way to treat breast cancer?
53:57Brain Mets, I think isn't that
53:58what the gamma knife is for?
53:59It's it's easy, right?
54:00You see a brain,
54:01but you just send the patient to the
54:02gamma knife and I would say, Oh no.
54:05It's not so easy.
54:06First of all,
54:07not everybody is a candidate for
54:08the gamma knife and gamma knife
54:10does work really quite well,
54:11but there are risks.
54:12Radiation, necrosis,
54:13as Doctor Chang will attest
54:15to is is a problem,
54:17and there's a lot of a lot of it that occurs.
54:19And when disease comes back after the game,
54:21and if it's even harder to take care
54:23of and then there are the patients for
54:25which gamma knife is unfortunately not an
54:27option that require whole brain radiation.
54:29And even with our fancy
54:30spinning of the beams,
54:31hippocampal sparing in the manting,
54:33it carries risks of neurotoxicity so.
54:37I, I don't think there's any question.
54:39If there was a way to reduce the
54:40need for radiation or at least lower
54:42the dose of radiation required,
54:43that would make a big benefit for
54:45patients with breast cancer and brain
54:48metastases and other tumors as well.
54:50So we have already started looking.
54:54Into this and we have a DoD grant
54:57to help us conduct this work.
54:59I think DX one is perfectly poised to
55:02lower the needed dose of radiation,
55:04and I think radiation is perfectly
55:06poised to help the X one get into
55:08the brain nuts because remember.
55:10DX1 is looking for and using DNA to get
55:13into the tumors so dose of radiation
55:15to increase tumor death and release
55:17DNA should recruit more DX1 to the brain.
55:20Mets and the bar graphs are
55:21shown on the bottom there.
55:22That's so that's exactly what we see.
55:25We see the most.
55:25DX one get in the brain.
55:26Mets in the mice that get treated
55:29concurrently with the radiation.
55:30And take it to the obvious extreme.
55:33Those mice that get DX1 with the
55:35radiation also have the best response
55:37we see the less number of metastases,
55:39so it's all working the way we expect.
55:41I understand it's all preclinical,
55:43it's in mice, but sometimes things
55:45start to come together and it's
55:47it's looking very promising to me.
55:49Obviously very biased and.
55:51Very grateful to Benedet and Marta
55:53and Caroline and who for all their
55:55work with this it's it's been tough
55:57but we've hung through here and it's
55:59it's going really quite well now.
56:02And again, can't do any of this
56:04without help from your friends.
56:06I had to cross out the word little and say
56:08a lot because jangle from neurosurgery has.
56:11Been great friend and
56:12colleague to me for years.
56:13Now he is an expert in all things related
56:15to the brain and brain tumors and ways
56:18to treat tumors in the brain and he's
56:20helped us figure out these models.
56:21He's very interested in the dxy antibody.
56:23We work together all the time,
56:25but he also does some other things.
56:26Believe it or not,
56:27a lot of other things.
56:29And just recently had a great
56:31paper in nature cell biology where
56:33he reported on a new effect of an
56:36LRRC 31 protein that significantly
56:38sensitizes breast cancer brain
56:40metastases to radiation therapy.
56:42So it's almost like the universe is.
56:44Telling us something because that
56:48LRRC 31 protein is great.
56:51They can't cross the blood brain barrier.
56:54I think I just spent the last 20 minutes
56:56talking about an antibody that can
56:58carry things across the blood brain barrier.
57:00So we got the Hanson lab and the
57:02shower lab working together and we
57:04can put them together and make a
57:06DX1 LRRC 31 fusion protein that I'm
57:09hopeful is going to be next in line
57:11to cross over the river through the
57:14woods and into the brain and increase
57:16sensitivity to radiation therapy.
57:18So hopefully that was of some
57:21interest to people listening here.
57:23I have a lot of people to thank.
57:25Obviously jeongbang Zhao.
57:27Great colleague.
57:29Of.
57:29Joe Contessa and Marta Bero in his lab
57:32have been incredibly helpful to us.
57:34My own lab.
57:35I'm extremely grateful to your efforts.
57:37I'm I'm usually at the gamma knife nowadays,
57:38so you probably are wondering
57:40where I've been here I am.
57:42If you're looking for me,
57:42it still looks like me,
57:43right?
57:44And the Yale Gamma knife team
57:46everybody is is great help.
57:48I'm appreciative to everybody but
57:50last anyone that knows me knows
57:52that I'm a fan of the Marvel movies
57:54and the best part about the Marvel
57:56movies is always there's one more
57:57cut scene at the very end, right?
57:59And I think if you.
58:00Listen very carefully and you
58:02look off into the horizon.
58:04You will see that there is a team
58:07coming assembled led by Pi Megan King.
58:10Getting the best breast cancer,
58:11brains researchers together to
58:13expand and improve DNA targeted
58:15therapies towards better breast
58:16cancer treatment with Pat Larusso,
58:18Megan Kingaby Patel, Ryan Jensen,
58:20myself and Zhangzhou,
58:21and we're very thrilled to have
58:23been notified.
58:24Just recently that we've received
58:25the YCC Team Challenge Award
58:27to conduct this work.
58:29With that I will take a breath
58:32and stop talking and answer any
58:34questions if there are.
58:36Well, I think we're a little short on time,
58:40so it's it's exactly 1 now.
58:43James that was great.
58:46I'm certainly extraordinarily supportive
58:47of anyone who wants to study breast cancer.
58:51Brain tests is an area that I've
58:53thought about a lot over the years,
58:55and I would agree with you 100% that it's
58:59an area that's perhaps the most in need,
59:04and potentially the most
59:05in need in breast cancer.
59:06Because we we we may well be able
59:08to eradicate disease elsewhere in
59:10virtually everyone in the next decade.
59:13But the brain is the hardest place,
59:14it seems so.
59:16With that I wanna thank both
59:19both of you for great talks.
59:21It's been a great grand rounds and James.
59:23Congratulations on the on
59:25the challenge award.
59:28Thank you so much, alright?
59:31I made thanks again bye bye.
59:35And thanks to our audience.