Richard Frisbee Memorial Lecture/Mechanisms of B-cell selection as therapeutic targets

Transcript

• 00:00First grand rounds of 2021 and once again
• 00:03continuing our our mission doing this.
• 00:06Whether it be in person or by zoom,
• 00:09we're sticking to the schedule
• 00:11and making sure that we we advance
• 00:14the mission of disseminating
• 00:16and knowledge through this form.
• 00:19Today is a really special occasion
• 00:22because it represents something I
• 00:24think we all look forward to the
• 00:27annual Frisbee lectureship and
• 00:28actually to begin the forum I I'm
• 00:31going to turn it over to Ed Snider,
• 00:34Doctor Schneider, as you know,
• 00:36is a professor of laboratory medicine.
• 00:41Can't send director for.
• 00:42For membership director of
• 00:43membership for the Cancer Center,
• 00:45Leader of blood banking.
• 00:47Certainly has done a lot of work
• 00:49over the years in that domain and
• 00:52has really been the Shepherd for this
• 00:54lectureship for the past 18 years.
• 00:56And I want to turn it over to Ed to
• 00:59share some perspectives and as well
• 01:02introduce members of the family.
• 01:05Thank you very much Charlie to pleasure to
• 01:08introduce the Frisbees for this lectureship.
• 01:10Rick and Chris Frisby, son Richie,
• 01:12developed leukemia as a young man that's
• 01:15a teenager and was the first bone marrow
• 01:18transplant done in the Connecticut.
• 01:19It was done by the late Jill Rappaport,
• 01:22who was running the program at the time.
• 01:26Richie did not survive first
• 01:28transplant failed and didn't
• 01:30survive to have a second transplant,
• 01:32which was to be his sister.
• 01:34And in honor of him, in his memory,
• 01:37the family rich Rick and Christine
• 01:40set up the Frisby Foundation.
• 01:42So in 1990 they've given millions of dollars
• 01:46in Cancer Research and cancer education.
• 01:49They established the first stem
• 01:51cell processing lab at Yale,
• 01:53New Haven long before Smilow had the
• 01:56first brick lay down the foundation and
• 01:59that was the precursor of the HCT lab,
• 02:03which is currently supporting SMILOW in
• 02:05through Department of Laboratory Medicine.
• 02:07Went by Diet Doctor Diane Kraus.
• 02:10This lectureship was established 18
• 02:12years ago and the current speaker
• 02:15doctor Marcus Motion is therefore
• 02:17the 18th speaker and we welcome him.
• 02:20And we would like to turn it over to
• 02:22Christine to do or say a few words.
• 02:24Of I'll be brief,
• 02:26I just wanted to really thank Yale Dr
• 02:28Mnuchin, and in particular at Snyder,
• 02:30who's been very close to us for many
• 02:33many years and been very supportive
• 02:35of the foundation and the work we do.
• 02:37And we love this lectureship. We don't.
• 02:40Foundations must smaller now.
• 02:41We don't do that many things,
• 02:43but this is one thing that we continue doing,
• 02:46and we're going to continue to fund
• 02:48this for years to come because we
• 02:50think it's just very rewarding.
• 02:52So thank you very much, Ed.
• 02:54Doctor Fox and Marcus as well at
• 02:58the Commission.
• 02:59Thank you Rick and Chris for all
• 03:01that you've done and for so much
• 03:03of the work that is being done for
• 03:06the patients that smile.
• 03:07Oh well,
• 03:07we can thank you for for setting
• 03:09the foundation for this,
• 03:11so I'll now turn it over to Doctor
• 03:13Fuchs to introduce our 18th.
• 03:15Frisbee lecturer Doctor Marcus machine.
• 03:18It thank you, Ann and Rick and Christine,
• 03:21thank you for your continued
• 03:24support of this important
• 03:25leadership over the past 18 years.
• 03:27I think this is really been
• 03:30a wonderful tradition.
• 03:31Becausw, what this lectureship
• 03:33has done is brought to Yale.
• 03:36Really rich cadre of innovators
• 03:39in developing understanding an
• 03:41new approaches to human logic,
• 03:44religion sees which I think is a fitting
• 03:48legacy and this year's annual Frisbee
• 03:51lecture is no exception to that.
• 03:54Impressive in August,
• 03:56List of lecturers.
• 03:58Doctor Marcus musician was previously
• 04:01the chair of the Department of
• 04:04Systems Biology and the Lee Professor.
• 04:07Oh, at the City of Hope Cancer Center,
• 04:11as well as the associate director
• 04:14of Basic Science an we were very
• 04:16privileged in the fall of 2020 to bring
• 04:20and recruit Doctor Mission to Yale.
• 04:23As our inaugural director for the
• 04:25Center of Molecular and Cellular
• 04:27Oncology at the Yale Cancer Center
• 04:30and Smilow Cancer Hospital as well
• 04:33as the author and Isabel Bunker,
• 04:35Professor of Medicine, focused in hematology.
• 04:38Doctor Mission,
• 04:39trained in hematology oncology.
• 04:41His work as his training was both
• 04:44in the biology of human like you
• 04:48put in season Immunobiology.
• 04:50And frankly,
• 04:51over the past decade or really longer,
• 04:54he has been a leading innovator
• 04:57in understanding the evolution of
• 05:00B cell malignancy's understanding
• 05:02biology in terms of delivering
• 05:05new approaches to drug discovery
• 05:07in these cancers,
• 05:09Ann and really now advancing that
• 05:11beyond in terms of the immunobiology
• 05:14immunotherapy that would be available
• 05:17for lymphomas and human logic legacies.
• 05:21Most notably narrow,
• 05:22even in terms of novel car T therapies,
• 05:26and I think all of that really
• 05:28speaks to in many respects what's so
• 05:31relevant for the Frisby lectureship
• 05:33Marcus for his many accomplishments
• 05:35and impressive publication record
• 05:37has been received countless awards.
• 05:40I I wouldn't want to take too much
• 05:43time annunciating all of them,
• 05:45but they include the NCI outstanding
• 05:48Investigator award.
• 05:49Howard, You scholar award, leukemia,
• 05:51Lymphoma Society scholar award.
• 05:53Welcome Trust scholar award.
• 05:55Among many other awards that
• 05:57recognize his accomplishment,
• 05:59an innovative and record of innovation and
• 06:02accomplishment across all of these cancers.
• 06:05So it's really a pleasure to introduce
• 06:08Marcus and an in many respects.
• 06:11Welcome to the faculty of Ovvio
• 06:15Cancer Center. Orchis, thank you.
• 06:20Thank you, I'm not sharing my screen.
• 06:46So first of all, I would like
• 06:48to thank the Frisby family and
• 06:51Charlie an ad for me being here.
• 06:55And since I came to your Cancer
• 06:58Center last for like I came to
• 07:01experience that many of us who
• 07:03are devoted to the cause of.
• 07:05Now titled can try to leukemia
• 07:08and leukemia and young tires.
• 07:10That legacy of Richard D.
• 07:13Frisbie has inspired many of us
• 07:15and and today I would like to text
• 07:19opportunity to present a few new
• 07:21findings from our lab that over
• 07:24the past recent years have led to
• 07:27a new concept that I hope will
• 07:30help us in the future to treat this
• 07:34disease is more efficiently than.
• 07:37We were able to do in the past.
• 07:39And many of our try to leukemias are
• 07:43actually derived from lymphocytes
• 07:44and is a leukemia.
• 07:46In fact,
• 07:47represents the most frequent type of
• 07:50cancer in children and young adults,
• 07:52and one potential reason for that
• 07:55is that B cells during the early
• 07:57development have to go through a
• 08:00series of genetic modifications
• 08:02in error combination class,
• 08:04switching hypermutation with the end
• 08:07goal for these cells protect us by.
• 08:10Generating Pi affinity antibodies
• 08:11and this is a cartoon here from
• 08:141905 drawn by Powell, Ellie.
• 08:16She was sitting here in his office
• 08:19and for those reasons B cells
• 08:22are an extremely high risk for
• 08:24malignant transformation.
• 08:26Actually,
• 08:27500 times higher than any other somatic cell,
• 08:30and for this reason and also becausw,
• 08:33humans can actually live without
• 08:35the lymphocytes for quite some time.
• 08:38And we developed a research program
• 08:41that is centered on specific
• 08:43vulnerabilities of this very cell type.
• 08:47So as cancer researchers,
• 08:48we are always looking for vulnerabilities,
• 08:51and in this case we're looking
• 08:54for vulnerabilities.
• 08:55That are intrinsically encoded in
• 08:57the nature of the sales making
• 08:59antibodies and are selected and
• 09:01ANVISA selection is a scene that
• 09:04that that I hope I will be able
• 09:06to present to you on the occasion
• 09:10of this lecture.
• 09:11And the reason is that just by the
• 09:14random nature of recombination,
• 09:17events of antibody encoding
• 09:19molecules vast majority,
• 09:20about 75% of oil newly generated
• 09:23visas are initially autoreactive,
• 09:24meaning that their directed against
• 09:27himself and in these cells have to
• 09:30be removed from the repertoire,
• 09:32and this means they have to be
• 09:35powerful mechanisms in place to
• 09:38normal development to destroy and
• 09:40delete yourself from the repertoire.
• 09:43And although arching theme for
• 09:45our research in recent years is,
• 09:47can we actually leverage these
• 09:49mechanisms that are indeed in the
• 09:52life and selection and development
• 09:54of normally lymphocytes for the
• 09:56treatment of Pisa, leukemia and lymphoma?
• 10:00So in.
• 10:00Fact,
• 10:01the principle of the cell selection
• 10:03is driven by signals from the B cell
• 10:06receptor or surface immunolabeling,
• 10:08and we like to think of this.
• 10:11Like I said, Goldilocks principle.
• 10:14Because only if the signal strength
• 10:17that is elicited from this unit
• 10:19here is just right,
• 10:21then this says receive a positive
• 10:23signal and proliferate and survive,
• 10:25and this is usually the case
• 10:28when we have a
• 10:29balance between activation signals.
• 10:33Namely, kinase and phosphatase is.
• 10:36That achieve this balance.
• 10:37So if the signal is too weak,
• 10:40for instance,
• 10:41gives the first parties are just wrong,
• 10:43or the receptor itself is not
• 10:46functional and he says die by neglect.
• 10:48Now we're focusing here on
• 10:50the other end of the scale,
• 10:52where the signals overwhelmingly strong,
• 10:54which is typically the case when this
• 10:56receptor he is engaged by self antigen,
• 10:59meaning that these receptors
• 11:01are out reactive.
• 11:02And these cells could give rise to autoimmune
• 11:06disease and have to be eliminated.
• 11:08And so this principle is not only
• 11:12relevant to normal B cell evelopment.
• 11:17In this cartoon here from
• 11:19a recent publication,
• 11:20shows that in transformed B cells and
• 11:23leukemia and lymphoma the signaling
• 11:25pathway downstream of the beasts
• 11:27are receptors engaged permutations.
• 11:29Every step of the way.
• 11:32And so today I would like to divide
• 11:34my talk in three areas where we gain
• 11:38information of how we can leverage
• 11:41selection for therapeutic benefits.
• 11:43One comes from inside that we glean
• 11:46from mutations and deletions and visa
• 11:49humorists such as leukemia and lymphoma.
• 11:51Then inside some clinical trials.
• 11:54So we collaborate with large
• 11:56clinical trial groups in the United
• 11:58States and internationally and
• 12:00look for predictors of clinical.
• 12:02Outcomes and what we can learn in
• 12:05terms of therapeutic targeting options.
• 12:08And then finally how these complicated
• 12:11oncogenic signaling pathways
• 12:13intersect and how we can leverage
• 12:16these interactions again to undermine
• 12:18oncogenic signaling in these diseases.
• 12:21So in the first part I'm going to
• 12:24talk about genetic lesions and what
• 12:26we have done here based on mutation
• 12:29data from cosmic and other sources
• 12:32assembled a set of more than 5 million
• 12:36somatic mutations in 39 different
• 12:38types of cancer and look at these
• 12:41mutations from the angle of whether
• 12:44the mutation introduce a replacement.
• 12:47Or effect according capacity of the
• 12:49gene or whether they are silent,
• 12:51meaning they are not selected for.
• 12:53And then in all these diseases we
• 12:56rank the mutations based on these
• 12:58replacement over silent ratios.
• 13:00And we do that in a way that ranks US
• 13:04based on the cell specific ratios.
• 13:08We end up with typically mutated genes,
• 13:10and these are widely known in B cell
• 13:13tumors like my D8820 and so forth,
• 13:15But was interesting to us that here
• 13:18at the top of the list they actually.
• 13:22Molecules in the PS3 kindness
• 13:24pass visit our frequently mutated
• 13:27throughout cancer,
• 13:28but are unexpectedly spirit were
• 13:30exempted from in B cell tumors,
• 13:33and this activating mutations
• 13:36of the PS3 kinase pathway.
• 13:39And P-10 and then ship one
• 13:41night in inventory.
• 13:42Phosphatases in this pathway.
• 13:43And this is the catalytic subunit
• 13:46of peers with kindness itself.
• 13:48So we studied this in multiple
• 13:50different directions,
• 13:51but I would like to focus your
• 13:54content because it was just such
• 13:57a subset of striking example and
• 13:59then as we know for a long time,
• 14:02pretend deletions and mutations widely
• 14:04occur throughout many cancer types,
• 14:06but in 925 cases of B cell image leukemia,
• 14:09we didn't find any of these mutations.
• 14:12Again, highlighting the
• 14:14specificity of this mechanism.
• 14:16And pretending opposers peers
• 14:19to kindness signaling by.
• 14:22I am targeting Pep 3 which is a
• 14:24central initiator of the PS3 kinase
• 14:27signaling pathway and I will come
• 14:29to that back later in my talk.
• 14:31And to study hyperactivation of the PS3
• 14:34kindness pathway by dilution of 10,
• 14:36we develop the mouse model in our lab
• 14:39based on conditional deletion of the
• 14:42P 10 gene in our leukemia model and
• 14:46unlike what we know and solid tumors,
• 14:49conditional deletion of P.
• 14:5010 result in Rapid City S of leukemia
• 14:54cells and if we change the sequence and
• 14:571st delete return a normal B cells and
• 15:01then bring in transforming Uncle gene.
• 15:03Then we basically compromise malignant
• 15:06transformation and then most importantly,
• 15:08when we wait for leukemia to
• 15:10establish and miles at bear these
• 15:12tumors and then delete speech engine,
• 15:15this leads to remission and the mice
• 15:18survive for indefinite periods of time.
• 15:21We also confirmed that the biochemistry
• 15:23or the premise here is correct because
• 15:26we actually do see increased activity,
• 15:29increased output of the PSC kindness
• 15:32pathway by increased phosphorylation of AKT.
• 15:35And most importantly,
• 15:36we used inhibitors that block the PS3
• 15:39kinase signaling pathways at multiple levels.
• 15:42Here,
• 15:42activation of PS3 kinese bicec using the
• 15:45sick kinase inhibitor and it's platinum.
• 15:48BKM 120 is a pan PSD.
• 15:51Kindness never turn easy.
• 15:53D 53 E 63 inhibits AKT and all
• 15:56three of them have in common.
• 15:59That's actually rescuer protect
• 16:01leukemia cells from sad as that
• 16:04would otherwise be in use.
• 16:06Dapon deletion of the P-10 phosphatase?
• 16:08So this doesn't mean that these
• 16:11compounds are counterproductive in
• 16:12leukemia because actually quite useful.
• 16:15Our interpretation of this
• 16:17unexpected result is that.
• 16:19I was introduction of those inhibitors
• 16:21restores signaling equilibrium again,
• 16:23the Goldilocks principle
• 16:24that these cells need,
• 16:26whereas deletion of the 10 alone here
• 16:29introduce a drastic perturbation
• 16:31which engages negative selection
• 16:32just as it does for the elimination
• 16:36of Hartree active users.
• 16:40We are hopeful that these ideas
• 16:43these concepts will eventually
• 16:45make their way into the clinic,
• 16:47and as an early indication that
• 16:49that might indeed be the case,
• 16:52I'm showing you two promising
• 16:54preclinical results in our lab,
• 16:56both based on small molecule inhibitors
• 16:58of these key fast watch cases, namely,
• 17:01ship one inhibited by 3A amino color stain,
• 17:04and then pretend which also is.
• 17:08Target if it was a small molecule
• 17:11inhibitor and both of them have
• 17:14desirable on target activity
• 17:16biochemically and importantly,
• 17:18their chief Disease Control or disease
• 17:21burden control for long periods of time,
• 17:24and she's,
• 17:25extension or prolongation of overall
• 17:28survival of mice that bear patient arrives
• 17:31in a graphs from B cell image leukemia cells.
• 17:36So we're hoping that this approach
• 17:39can be developed further in and
• 17:41that some of these compounds will
• 17:43make it into the clinical arena.
• 17:46Now the central premise of this
• 17:49idea is that this is a mechanism.
• 17:52Negative selection.
• 17:53Removal of art reactive says that is
• 17:56uniquely important in B lymphocytes
• 17:58and to test this premise we performed
• 18:01a reprogramming experience.
• 18:06And in which we should use the VPI flower,
• 18:09just a transcription factor that can
• 18:12transform besides into macrophages.
• 18:13And this is shown here besides expressed in
• 18:1719 and this marker here is lost overtime
• 18:20after induction and in favor of Mach one,
• 18:23which is a macrophage marker.
• 18:26And indeed, after sometimes you say it,
• 18:28start to crawl around on the bottom
• 18:31LCS dishes and like macrophages and
• 18:33can even phagocytose and importantly.
• 18:36Coming back to our hypothesis,
• 18:38if you genetically delete P.
• 18:4010, Even though genetically identical.
• 18:43The reprogramming from B to Milo to be 2.
• 18:47Macrophage fade almost entirely
• 18:49removes the sensitivity of these cells
• 18:52to removal of source phosphatases,
• 18:54which makes sense because fact
• 18:56macrophages don't make autoantibodies,
• 18:58there's no need for macrophages to be
• 19:01negatively selected as beast cells are.
• 19:03So this gives us confidence that this is
• 19:07a real mechanism that is reflective of
• 19:10the nature of the immune system to purge.
• 19:14Attractive sales and that is possible to
• 19:17selectively target this vulnerability
• 19:19in B cell tumors.
• 19:21Now in this work was done by Gen.
• 19:24John Shannon, our lab,
• 19:26and when he worried that this is not
• 19:28just counterintuitive because you
• 19:30essentially doing the opposite from what
• 19:33everyone else is doing in this field,
• 19:35namely by instead of inhibiting kinases.
• 19:37VR Pro activating kinases.
• 19:39But most importantly,
• 19:40what was worried about what happens
• 19:42if he hyper activate kinases for
• 19:44long periods of time,
• 19:45because that in itself could be dangerous,
• 19:48so he did an experiment to figure out
• 19:51what is the shortest period of time.
• 19:54Home to commit Visa is to say this and he
• 19:57did this with an engineered hyperactive.
• 20:00Formosa sick kinase.
• 20:02Labeled here as GFP.
• 20:04If you bring in this hyper active
• 20:07kinase in the presence of stickiness,
• 20:09inhibitors of GFP labeled cells
• 20:11remain constant,
• 20:12'cause there's no hyperactivation
• 20:13of the pathway.
• 20:15Now,
• 20:15if we wash out the inhibitor
• 20:17cells as expected, rapidly die,
• 20:19and he found that if there's
• 20:21a lapse of just three hours,
• 20:24so removal of simulator for three hours
• 20:26and then adding it right back that
• 20:29already is sufficient to commit the sales,
• 20:32that will be our goal going forward too.
• 20:35Target is short.
• 20:37Strong exposure to hyperactivation
• 20:39probably was click or only dated compounds
• 20:42that have a short plasma half life.
• 20:45Now in the second part of my talk,
• 20:48I'm going to give you 2 examples of how
• 20:51we can learn from information within
• 20:54clinical trials and gene expression,
• 20:57annotation related to outcome.
• 20:59So what approach is based on
• 21:02microarray data that we obtain?
• 21:04Collaboration was a clinical study.
• 21:06Groups and much of this works
• 21:10also publicly available.
• 21:11And for each of these micro area
• 21:13probe sets that measure expression
• 21:15of individual transcripts,
• 21:17we divide the patient courts and
• 21:19the two groups based on higher than
• 21:21median versus lower than median
• 21:23expression in these clinical trials.
• 21:25And then we asked the question,
• 21:28is there a difference between those two
• 21:30groups in terms of clinical outcome?
• 21:33And if the outcome is more favor
• 21:35we have here
• 21:37a blue annotation and its outcome is
• 21:39more poor, shorter overall survival.
• 21:41For instance, we have a red annotation
• 21:44and if the group this heat map based
• 21:46on the site specific annotations we
• 21:49come up with a list of genes that
• 21:52became interesting to us and he at
• 21:54the very top is 1 molecule that I'm
• 21:57going to spend the next couple minutes
• 21:59on the L2 receptor Alpha chain,
• 22:02also known as C25.
• 22:06So that was a bit unexpected.
• 22:09Becausw CD 25 is known as one of
• 22:12the three chains of the L2 receptor.
• 22:15Anna typically pairs with the better
• 22:17chain in the gamma chain to form a
• 22:20trimeric receptor, and this was step
• 22:22was active on T cells and in cases.
• 22:26And it's also important for
• 22:28formation of regulatory T cells or T.
• 22:31Rex, and therefore therefore
• 22:33important to prevent autoimmunity.
• 22:34Again, important to be self selection.
• 22:37And here I'm showing you that.
• 22:41Then if you look at individual visa diseases,
• 22:45pediatric B cell, leukemia,
• 22:47CLL, Podiatry, pizza, leukemia,
• 22:49mantle cell lymphoma,
• 22:50we see consistent pattern that
• 22:53the lower half of expression
• 22:56is related to better outcome.
• 22:58The top half towards outcome.
• 23:02The other reason we became interested
• 23:04is that if you bring in Uncle Gene,
• 23:07said drive, leukemia,
• 23:08lymphoma like these are able or LMP 2A.
• 23:10This leads to upregulation
• 23:12of C25 on the cell surface.
• 23:15And also it seems to play a role
• 23:17in resale developmented save.
• 23:19So he profile here see 25 M on A levels.
• 23:24Over the course of the sale
• 23:26evelopment we find here is striking
• 23:28peak and the so called Faction D.
• 23:31And that's interesting,
• 23:32because if you look at C25 knockout mice,
• 23:34which we did in our lab.
• 23:37Compare this to the wild type
• 23:39animals and look at these fractions
• 23:41we see here is fraction D.
• 23:43But distractions entirely missing in the
• 23:46knockout mice and can also see this here.
• 23:48This is a defect here.
• 23:50We still don't know what this actually means,
• 23:53but we also find that later in
• 23:56development he says actually are
• 23:58over represented in fraction F.
• 24:00So our initial hypothesis was this might
• 24:05reflect previously unrecognized role
• 24:08of IL two signaling in B cells and.
• 24:11I mean,
• 24:12so we repeated this experiment with
• 24:15mice that have intact City 25,
• 24:17but are lacking the L2 cytokine.
• 24:20But contrary to our hypothesis fraction,
• 24:22D&F are just fine and be so developmen
• 24:26is completely unperturbed in these mice.
• 24:29And I'll see you in this proximity
• 24:32ligation analysis,
• 24:32we find that C25 does actually
• 24:34not bind to any of those other
• 24:37change of the L2 receptor and does
• 24:39not respond to iron tools.
• 24:41So it's not true that C.
• 24:4325 is in any way related to I
• 24:46L2 signaling and visas instead.
• 24:49We found in our proximity ligation assay,
• 24:52said City 25 associate itself.
• 24:54With a signal image chain of
• 24:57the B cell receptor,
• 24:58which again is responsible for the
• 25:01Goldilocks principle to keep intact
• 25:03and equilibrium and intermediate
• 25:04ram of signaling intensity.
• 25:06And that's the case in resting B
• 25:09cells where prices are even more so
• 25:11the case after the visa receptor was
• 25:14stimulated with an anti IG M antibody.
• 25:17This is actually ongoing work in our lab,
• 25:20by Jay,
• 25:21wrongly with a research scientist
• 25:24in my group.
• 25:25And what he found is actually their CIA.
• 25:2825 negatively regulates B cell
• 25:31activation and in the absence of CD 25.
• 25:34Miles,
• 25:35develop spontaneous germinal center,
• 25:37so even without any immunization,
• 25:39these B cells are autoreactive.
• 25:42Their escape negative selection
• 25:44and therefore more attractive,
• 25:47spontaneous germinal centers
• 25:49that are antigen independent.
• 25:51The other observation here was that if
• 25:55Jerome deleted 325 in human lymphoma sales,
• 25:58they undergo a particular pattern
• 26:01of autonomous calcium signaling.
• 26:03They have autonomous activation.
• 26:05Do sales are proliferating very fast,
• 26:08but also for short half life and I quickly,
• 26:12which is reflected here by
• 26:14expression of PG restore parenting.
• 26:17These cells are.
• 26:18Just easily exhausted and
• 26:20in that competitive fitness,
• 26:23so we confirmed this here in a
• 26:27leukemia model whereby we transformed.
• 26:3025 mouse cells with the flux see 25
• 26:34every year and then after activation
• 26:37of query is illusia CD 25 expression
• 26:40on the surface and then soon after
• 26:44this says disappear from culture.
• 26:46They failed to form any colonies that
• 26:50cannot initiate leukemia and mice
• 26:52that bears also leukemias recover and
• 26:56survive for indefinite periods of time.
• 26:58Now, Interestingly,
• 26:59and that's coming back to
• 27:01signaling feedback control,
• 27:03we found that upon collisional
• 27:05City 25 in a similar way like
• 27:08deletion of P-10 and ship one,
• 27:11we see that the balance of ether
• 27:14receptor signaling strength.
• 27:16It's lost cause we have hyper
• 27:19activation of kinase substrates
• 27:21downstream of the visa receptor,
• 27:23including sick and then loss of phosphatase
• 27:27activity markers for P-10 and ship one.
• 27:31So we think those phenomena
• 27:33might actually be related,
• 27:34but CD 25 plays a role in maintaining
• 27:37the Goldilocks principle by
• 27:40regulating kinases and phosphatases.
• 27:42Now,
• 27:42how is this possible?
• 27:44So the tail of CD 25 is very short
• 27:47here and it's just 13 amino acids and.
• 27:51So we looked at what City 25 might bind to.
• 27:55How does it interact with the
• 27:58cytoplasmic tail and as a negative
• 28:00control and using this for a
• 28:02lot of different experiments,
• 28:05we introduce a mutation of the
• 28:07central Seren residue which
• 28:09destroys the main protein kinase.
• 28:12He better consensus motive.
• 28:14And here we are using a bio ID
• 28:18approach which is based on fusions
• 28:21between the CD 25 tail and puree,
• 28:24which is a bacterial biotin ligase
• 28:27which attaches bio tends to approximate
• 28:30protein space on the mound and the proximity.
• 28:34And I,
• 28:35as expected,
• 28:36we found that two phosphatases
• 28:38ship one and PTPN 6 here.
• 28:41Are in proximity of the tail of
• 28:43CD 25 and this is not the case.
• 28:46The tail here is mutated.
• 28:49And this is also confirmed here in
• 28:52a more traditional experiment based
• 28:54on pull down and quiet peace or
• 28:57ship 1P-10 and PTPN 6 can or bind.
• 29:00But binding this weekend or entirely
• 29:03lost when the stay here is mutated.
• 29:06So in terms of function,
• 29:08this could be confirmed that indeed see
• 29:1125 functioned as a powerful negative
• 29:13regulator of signaling strength.
• 29:15So if he abusively activate while
• 29:17types unify, we can block the kite.
• 29:20Some signal here that would
• 29:22otherwise be elicited.
• 29:23So he expresses the 25
• 29:25and it's wild type form.
• 29:27The signal is delayed and
• 29:29almost entirely lost if they
• 29:31express the mutant.
• 29:32This depression can still be seen,
• 29:35but it's much less compared
• 29:37to the wild type form.
• 29:40And then in terms of leukemia,
• 29:42survival and growth be used here,
• 29:45cameras between the
• 29:47extracellular part of CD 19,
• 29:49which is a B cell specific
• 29:52transmembrane protein and tale of
• 29:55225 users via type or is mutant
• 29:57and wild type form can rescue.
• 30:00Survival of leukemia cells but
• 30:03seven 268 a mutant cannots again
• 30:06showing that ability to recruit
• 30:09phosphatases to negatively
• 30:10regulate signaling is important for
• 30:13survival of these leukemia cells.
• 30:16So we have modeled the interaction
• 30:19between these molecules and came up
• 30:21with the structural model for this,
• 30:24which has rank one PKC better
• 30:27scaffold at the center.
• 30:29Wrapped around by PKC better
• 30:32and this interaction,
• 30:33he has facilitated by the C25
• 30:36tear which insert itself here.
• 30:38So overall we think that he says.
• 30:42Activate CD 25 downstream of the
• 30:44visa receptor via sick because
• 30:46he better phosphorylation of C25,
• 30:49which then forms a complex with Raekwon
• 30:52to recruit first parties here to surface.
• 30:55Which then again provide negative
• 30:57feedback control so it's like a
• 31:00circle that goes back to maintain
• 31:03equilibrium Goldilocks principle
• 31:05again for the survival of the cells.
• 31:07So our conclusion is that we think that
• 31:11negative selection can be leveraged indeed.
• 31:14For potential therapeutic benefits of.
• 31:17He said leukemia and lymphoma is
• 31:20avoided because of phosphatases
• 31:22hyperactivation of sick or
• 31:24interference proceeding 25.
• 31:26Feedback control.
• 31:27So the goal here would be to.
• 31:30Push says that I had the upper limit
• 31:33there or it transformed the after a
• 31:36powerful activation signal over the
• 31:38edge by removing feedback control and
• 31:41balance which will trigger negative
• 31:43selection of what looks like at the
• 31:47level of signaling autoreactive B cells.
• 31:51Um?
• 31:51Coming back to our.
• 31:54Database which we find us a very rich
• 31:58resource for new ideas and concepts.
• 32:01You found another interesting
• 32:02outcome predictor that I would
• 32:04like to introduce to you with the.
• 32:06It's a recent publication that just
• 32:09came out a couple weeks ago and that is
• 32:12focused here on a molecule called IIT M3.
• 32:15And it's interfering inducible
• 32:18transmembrane protein.
• 32:19And as I showed you for C25
• 32:24to smaller cure is.
• 32:26An outcome predictor in various.
• 32:28He said leukemia and lymphoma
• 32:31subtypes and it's known for long time
• 32:35initially was found as a specifying
• 32:38molecules for primordial germ cells.
• 32:41And then more recently it was
• 32:43found as an antiviral protein that
• 32:45can restrict viral replication.
• 32:47He is shown HIV,
• 32:48but more recent data shows it also
• 32:51important for the restriction of
• 32:53coronavirus and many other viruses,
• 32:55and what was important to us is that
• 32:58is actually used as a diagnostic
• 33:00tool for pediatric leukemia to find
• 33:03patients that are at high risk.
• 33:05So it's one probe set on a low density
• 33:09array to identify patients at high risk.
• 33:12So we started the function of
• 33:14items three in a genetic mouse
• 33:17model and found actually.
• 33:19Happy says surprisingly,
• 33:20that are lacking this interferon
• 33:23inducible transmembrane
• 33:24protein. I have a defect
• 33:26in PS3 kind of signaling.
• 33:28And they are prone to
• 33:30cell death as shown by P.
• 33:3153 activation and loss of PCL too.
• 33:34And importantly, these cells actually
• 33:37cannot be properly activated to
• 33:39undergo affinity maturation.
• 33:41So here is PNA is a German center marker
• 33:44which is a throwback for affinity maturation.
• 33:48In in in, in visa here.
• 33:51So if he smiles are immunized and
• 33:55nicely form germinal centres but if.
• 33:58He says that by Adoptively
• 34:00transferred are lacking item 3.
• 34:03The amount of general centers
• 34:04or German centre visa is and
• 34:07subsequent affinity maturation.
• 34:08Is this drastically reduced?
• 34:11And the same is true in leukemia,
• 34:14so I have items with efficient leukemia.
• 34:16Says cannot form colonies.
• 34:18If Transformers disable or N Ross.
• 34:21There also lacks ability
• 34:23to initiate leukemia,
• 34:24and those two models.
• 34:26And they have a similar phenotype as I
• 34:29showed you in normally says in terms of
• 34:32lack of peers with kindness signaling.
• 34:35Survival and strong expression
• 34:39of death related or checkpoint
• 34:43related molecules like P53.
• 34:46So in terms of structure and mechanism,
• 34:49we were able to figure out how I've
• 34:52item three is regulated in Indy.
• 34:54Lymphocytes is actually very short
• 34:56protein or 433 amino acids in length
• 34:59and we found that it can actually
• 35:02insert itself into the cell membrane.
• 35:05And this happens when I'm downstream.
• 35:08Also visa receptor.
• 35:10Lynn,
• 35:10another sack family kinases phosphorylate.
• 35:14I've item three at this tyrosine 20s
• 35:16that's really a central tiersen,
• 35:18which leads to recruitment to
• 35:20the cell membrane,
• 35:21and then it can easily interact
• 35:24with the visa receptor or it
• 35:27becomes internalised again.
• 35:29So for this reason we studied this
• 35:31mutation here and actually found that
• 35:33it can function as an Uncle gene.
• 35:36So when we introduce us for cinematic
• 35:39form of white 20 which mimics the
• 35:42confirmation that is always was formulated.
• 35:44Into a mouse strain that
• 35:47carries transgenic BCR ABL,
• 35:48which has a very long latency to disease.
• 35:52We actually found that this leads
• 35:55to increased formation of colonies.
• 35:57Increased PS3 kinase activity and
• 36:00also increased activity of the
• 36:03visa receptor signaling pathway.
• 36:06And.
• 36:08And so structurally,
• 36:10we could show the poisoner interactome
• 36:12analysis that this form of items
• 36:15three intersects with multiple central
• 36:17components of both the PS3 kinase
• 36:19and visa receptor signaling pathway.
• 36:21And there's also shown here by these red
• 36:24dots in this proximity ligation assay,
• 36:27where if I can three molecules come
• 36:30in close proximity. And he says.
• 36:34Now the structural basis for data
• 36:37set and that was very surprising to
• 36:39us that I've item 3 can directly
• 36:42bind to PIP 3 which is initiating
• 36:45phospholipid and lipid rafts to
• 36:48initiate PS3 kinase signaling.
• 36:50And that is unexpected because this
• 36:53interaction is usually mediated by a
• 36:55so-called pH domain in larger proteins.
• 36:57But I've item three is such a short
• 37:00party in that has no resemblance
• 37:03of the pH domain,
• 37:04so we were looking here for and you
• 37:08structure basis of houses interaction
• 37:10could happen in the absence of a
• 37:13pH domain. And we looked at the
• 37:16conserved intracellular loop of
• 37:18five items free that that is used
• 37:21to insert into the cell membrane,
• 37:24and in doing so we found a cluster of.
• 37:30Five basic amino acids and of particular
• 37:33interest is this bracket here would
• 37:36call it between lysing 83 and license
• 37:39104 and even though they are 21
• 37:42amino acids apart from each other,
• 37:44they come very close here in
• 37:46the structure analysis,
• 37:48and they're basically former
• 37:50clamp to directly interact here.
• 37:52With this pit three molecule.
• 37:55So by mutation analysis we were able to
• 37:58show that the whites are morally cure.
• 38:01Was this bracket of lice and
• 38:0483 and license 104 intact?
• 38:06Is a powerful initiator of PSV kindness
• 38:09and peace a receptor signaling?
• 38:12But when these two amino acids
• 38:14here are mutated through the
• 38:16brackets along the active,
• 38:17the entire Lee loses ability.
• 38:20So that's something that became
• 38:22really interested in that we hope
• 38:24to pursue further in collaboration
• 38:26with our colleagues at.
• 38:27Yeah,
• 38:28like a new way of how proteins
• 38:30can make contact with Pepsi to
• 38:32initiate PSV kind of signaling
• 38:34in normal and Uncle Genic Lee.
• 38:36Transform B says so.
• 38:38How model is that in the
• 38:40absence of five items,
• 38:41we and normal cells also modeling and says.
• 38:45The molecules that initiate PSU
• 38:47kind of signaling are scattered
• 38:50throughout the cell membrane.
• 38:52Only five items,
• 38:53three is there acting as a molecular
• 38:56glues are drawn together and form a
• 38:58tight complex to initiate signaling.
• 39:01I'm not coming to the last part of my talk,
• 39:05which was quite surprising
• 39:07to some of us and Mr.
• 39:10Looking for ways to translate that knowledge.
• 39:13Looking for houses can be
• 39:15exploited therapeutically,
• 39:16but it essentially starts from the
• 39:18question of how do oncogenic pathways,
• 39:21once activated by mutations?
• 39:23How do they interact and?
• 39:27Becoming. Part of an orchestrated move
• 39:30that is to malignant transformation.
• 39:33And this idea is based on a concept
• 39:37that was formulated and long
• 39:40time ago by fear and Vogelstein.
• 39:44Here's Arconic concept of Mikey
• 39:46step malignant transformation by
• 39:49sequential acquisition of Driver
• 39:51Uncle Gene set together, then form
• 39:54the development of colorectal cancer.
• 39:56So the question here is, is this.
• 40:00Same in visa is do we.
• 40:03Is it true that acquisition of addition
• 40:05mutations lead to more malignant phenotypes?
• 40:07And how do these oncogenic
• 40:10pathways interact with each other?
• 40:12So to answer this question,
• 40:13we formed a collaboration with Children
• 40:16psychology Group and Saint Jude.
• 40:18And studied.
• 40:20Mutation data from one 1148
• 40:24cases of ecel image PLA.
• 40:27And what we did first was what you
• 40:30would call a mapping analysis of
• 40:33affinity versus repulsion of pathways.
• 40:35So basically asking the question.
• 40:37So activating lesions in one pathway.
• 40:41Are they?
• 40:42Do they have affinity to activation of?
• 40:48Hidden in a different pathway?
• 40:50Or is there like relationships of mutual
• 40:52exclusivity and we found a number of
• 40:54interactions that we are still working on?
• 40:57But one was really striking to
• 40:58us and it's an interaction of
• 41:01repulsion of mutual exclusivity,
• 41:03as shown here in this cartoon.
• 41:05And that involves a stat 5 pathway.
• 41:08The Jack Stat 5 Path pathway and IIRC
• 41:11home up kinase signaling pathway.
• 41:15And here I'm showing you the
• 41:18result based on these 1148 cases.
• 41:21Many of them have shown here in Green
• 41:24Spot 5 activating lesions forming
• 41:26one large cluster up here and then.
• 41:29Here's another cluster,
• 41:30but these are activating lesions and only
• 41:34in 35 cases which is just feed the spend.
• 41:36We found activation of both pathways,
• 41:39which which is much lower than than random.
• 41:43Also, when we look at individual
• 41:45cases and look at phosphorylation of
• 41:47Erk or phosphorylation of stat five,
• 41:50we have a clear cut negative or
• 41:53inverse relationship between them
• 41:54and you can also see here I have to
• 41:57level off Western blot that that
• 41:59you have either force relation or
• 42:01step file for false for work and
• 42:03this leads to different profiles
• 42:05in terms of correct sensitivity so
• 42:07traumatic Nip is American emitter.
• 42:09It was in the herb signaling
• 42:11pathway that effects on these.
• 42:13Leukemias hear worse porn atnip effects,
• 42:18mainly,
• 42:18the stat 5 signaling pathway which
• 42:21is affecting those leukemias here.
• 42:24And I'm.
• 42:26So we became interested in this small
• 42:28minority of cases in which we have
• 42:31indication of activation of both pathways,
• 42:34even though they seem to
• 42:36be mutually exclusive.
• 42:37So wanted to know who they occur in
• 42:40the same say or how does this work?
• 42:44And to answer this question,
• 42:46or we developed in our lap a single
• 42:48self also protein analysis that allows
• 42:51us to interrogate her phosphorylation
• 42:53of STAT 5 and phosphorylation of Erk.
• 42:56Concurrently in single cells,
• 42:58and this is here based on the gel
• 43:02matrix where we can deposit 6400 cells,
• 43:06single cells and then look at STAT
• 43:095 and workforce relations events
• 43:12individually and this year or
• 43:15four patient arrived.
• 43:17Cases where we looked at individual
• 43:19sales and were then actually able
• 43:22to determine that even though for
• 43:24all those four cases we get to dual
• 43:27signal by Western blot, if he.
• 43:30Use our single cell for supporting analysis.
• 43:33We see that these are actually two
• 43:35competing clones, 1 colonial start,
• 43:37five Zelda clone, here's org,
• 43:39and that goes for all four cases and we
• 43:42don't see any double expressing cells.
• 43:44So our conclusion is at least are
• 43:46actually rare by colonial diseases,
• 43:48in which two clones are competing
• 43:52against each other.
• 43:53Then we asked what is the reason for that?
• 43:57So what is the underlying mechanism
• 43:59that these two pathways just can't go
• 44:02together and to address this question,
• 44:04we actually voiced it.
• 44:06The alternative pathway on the leukemia
• 44:08said are driven by the other pathway,
• 44:10meaning that here is a visa able or
• 44:13start five driven leukemia then was.
• 44:16And Ross, when Ross driven leukemia was VCR
• 44:19able and use different models for that.
• 44:22And here, this colony forming assay
• 44:25shows if you have one single driver,
• 44:27either in the rason start.
• 44:29Five pathways is dramatically
• 44:31increases number of colonies,
• 44:33but if we have posed together,
• 44:35we basically lose or colony
• 44:37formation capability.
• 44:38The same holds true for growth,
• 44:41so single driver nicely lead to outgrows,
• 44:45but combination of Bosa suppressive.
• 44:48What was really surprising to us?
• 44:51This actually,
• 44:51that if we use genetic ablation of
• 44:54the diverging or alternative pathway,
• 44:56even though we basically
• 44:58remove an Uncle Genic driver,
• 45:00this actually Slack celebration of
• 45:02leukemia initiation in this model.
• 45:04So in this case we have here a
• 45:07visa able or start five driven
• 45:10leukemia and we remove perk. This.
• 45:14Accelerates development of leukemia.
• 45:16Likewise in a chaos driven leukemia.
• 45:20Removal of stat 5.
• 45:23Come initiates faster development
• 45:26of looking more Genesis.
• 45:29And biochemically,
• 45:29we were able to recover too late.
• 45:32This was small molecule inhibitors
• 45:35that Rametta Nathan MacKinnon bitter.
• 45:37Distinguishes her kindness
• 45:39activity as expected.
• 45:41But it also induces phosphorylation
• 45:43of STAT 5, and rocks.
• 45:45Litten appears opposite effect.
• 45:47It distinguishes start 5,
• 45:49but increases per activity.
• 45:51And so to end here,
• 45:54this final chapter of my talk,
• 45:57we came across agonists that
• 46:00we use for pharmacological
• 46:02reactivation of diverging pathways.
• 46:04So in this case BC I hear this,
• 46:08I 215 is so powerful activator, IIRC agonist.
• 46:12Strongly activates falsework at the
• 46:16expense of start 5 and then DPH here.
• 46:20Is the stat 5 agonist drive start
• 46:23five phosphorylation but at the
• 46:25expense of work and this has
• 46:27interesting activity on the clonal
• 46:29dynamics of these leukemias in vivo.
• 46:32So start five leukemia can be converted into,
• 46:35IIRC, leukemia, and most importantly,
• 46:37if you combine these pathway
• 46:39agonist was conventional treatment.
• 46:41So, for instance,
• 46:42here on Earth inhibitor with
• 46:44a Step 5 agonist,
• 46:46we achieve a dramatic
• 46:48prolongation of overall survival.
• 46:50So.
• 46:52The final conclusion here is that we
• 46:55propose that diversity of signaling input.
• 46:58Is actually an important barrier
• 47:01of malignant transformation and
• 47:03centralization and convergence
• 47:04onto one single pathway.
• 47:06Inactivation of all the other pathways.
• 47:10Is an early and critical step
• 47:12of malignant transformation,
• 47:13and if we achieve.
• 47:15To reinstate at the very signaling
• 47:18environment.
• 47:19That would resemble the interactions
• 47:22of normal cells were sent by him
• 47:25and which have multiple receptors.
• 47:27Multiple cues from the environment.
• 47:30So we're proposing a strategy
• 47:32of pharmacological reactivation
• 47:33that would restore their diverse
• 47:36signaling environment.
• 47:37And we hope that this approach can also
• 47:40be leveraged to overcome convention
• 47:42mechanisms of black resistance.
• 47:44So here we have passed their
• 47:46convergence minimal.
• 47:47What we called friction and permissive
• 47:49environment for transformation.
• 47:50But here if you have divergent pathways,
• 47:53we actually do have some friction and
• 47:56create a non permissive environment.
• 47:59And with that I would like to thank a
• 48:02number of collaborators at a dinner,
• 48:04Farber at yeah.
• 48:06And would like to acknowledge
• 48:08particular 2 senior members of my lap.
• 48:10Linda Shannon and Jerome Lee,
• 48:12who did most of the conceptual
• 48:14innovation of this work.
• 48:15Thank you very much.
• 48:19Marcus that's amazing. Collection
• 48:22of studies and it is remarkable how
• 48:27complex and somewhat almost counter
• 48:30intuitive alot of these pathways are
• 48:34in B cells and be some legacies.
• 48:38And it's obviously a delicate balance.
• 48:39And let me ask you, 'cause you?
• 48:42You've identified a number of pathways.
• 48:44That I guess are principally
• 48:47designed for B cell elimination.
• 48:49That you could leverage.
• 48:51How would you potentially
• 48:53target those pathways?
• 48:55Or could you target those pathways
• 48:57in conjunction with the growing
• 48:59armamentarium of available
• 49:00therapies for be similar?
• 49:02Concedes that are now in practice?
• 49:04Or could you,
• 49:05could you leverage that combination?
• 49:09Right in terms of potential for
• 49:11translation and how we would leverage
• 49:14hyperactivation of Visa receptor
• 49:16signaling to engage negative selection,
• 49:19and I think we have two options, one is.
• 49:24Already available, but less attractive,
• 49:27that would be April inhibition or
• 49:30phosphatases like Ship One and P-10.
• 49:33I'm attractive cause in the field
• 49:36phosphatase innovation raises a lot
• 49:39of questions about specificity.
• 49:41An alternative approach is one that
• 49:43Mark Lemon and I have discussed and that
• 49:47would be to generate a direct agonist
• 49:50of picky kinase like sick or BDK.
• 49:53Something fairly upstream in the
• 49:56visa receptor signaling cascade.
• 49:58So actually yesterday.
• 50:01I was at at Mark CBI weekly meeting and
• 50:03and there was some encouraging feedback
• 50:05that that might actually be feasable.
• 50:08So that's an approach that I would
• 50:10definitely like to pursue in the
• 50:12future to develop a direct hyper
• 50:14agonist or one of these key kindness.
• 50:19Excellent, thank you so and
• 50:21people should feel free to submit
• 50:23questions on the on the chat box.
• 50:26Dying cross as a question.
• 50:28I don't know if you can see it.
• 50:32Marcus, I'll just read it.
• 50:34PLA suggested that CD 25 C
• 50:3679 eight colocalization is
• 50:37predominantly intracellular.
• 50:39What do you think that this indicates?
• 50:43Great questions so.
• 50:47We have done an experiment with.
• 50:50WGA, which is a surface marker and
• 50:53we we actually find colocalization
• 50:55of about 60 to 70% of those
• 50:58interactions with the CD79A and B.
• 51:01So I don't think it's predominantly,
• 51:04but the question remains,
• 51:05there's still a significant amount of
• 51:08interactions that are was in the sale.
• 51:10And you thought, indeed,
• 51:12that is very strange,
• 51:13because why would the visa receptor
• 51:16be internalized or somewhat was in
• 51:19this area associated with CD 25?
• 51:21And so we can answer this fully.
• 51:24But in light of the recent study
• 51:27by whose daughter was published
• 51:29in nature two years ago,
• 51:31where he actually shows that in malignant
• 51:33lymphoma B cell receptor signaling
• 51:36complexes form of endosomal membranes,
• 51:38his Tinder sale.
• 51:39We think that exact same
• 51:41thing might happen here.
• 51:43You don't know that,
• 51:44but that's that would be our explanation,
• 51:46so I don't think it's predominant.
• 51:48But I agree with Diane that let
• 51:50me see intracellular complexes,
• 51:52and we think they're an endo zones.
• 51:55Thank you and Diana's second question,
• 51:58which is CD 25 seems to
• 52:00prevent autoreactivity.
• 52:01Do you think this is related
• 52:04to CD-25 CD 79 interaction?
• 52:07Does it? Does C25 interact with
• 52:11surface immuno globulin's? I'm.
• 52:14So. That's actually a question
• 52:17that I asked Eric Metra,
• 52:19who's my collaborator in the
• 52:21field of autoimmune diseases,
• 52:22so Eric told us at 3:25,
• 52:24and that's known by work from
• 52:27from his group and also others.
• 52:29It is crucial to maintain
• 52:32central visa tolerance so that
• 52:34molecule is not there anymore.
• 52:37Then central tolerance mechanisms don't work.
• 52:40I think our signaling studies
• 52:42just about to clarify how this.
• 52:44Actually,
• 52:44you know what this mechanism of that?
• 52:47I think the link or how C25 interferes
• 52:49with Visa receptor signaling is not
• 52:52known in our paper is not published yet,
• 52:55so we're still working on that.
• 52:57Honey.
• 52:58And I I do think,
• 53:00actually,
• 53:00that it doesn't act in
• 53:01service in the global India.
• 53:05And again, people should
• 53:07submit their questions online.
• 53:08Marcus, I mean, could you ever
• 53:11conceive of you know you identify a?
• 53:14Be so malignancy that's driven by
• 53:16jackstadt and you would give them,
• 53:18IIRC activator.
• 53:18You know I obviously we focus on how
• 53:21to inhibit the pathway in cancer,
• 53:23but is that something you could
• 53:25conceive as a therapeutic approach?
• 53:29If I may quote back on my slides
• 53:31because there's one that I want
• 53:33to show you. This is study.
• 53:37Yeah, this study heals. Come by Veronica.
• 53:40Sex is group so that did you
• 53:43want to share your slide or? Yes,
• 53:46I'm going to show this slide again.
• 53:54OK, so I I hope I'm I
• 53:57got the question correct.
• 53:59This is what I I would like to refer.
• 54:02It's a study by the only car sex is cool.
• 54:07That it's a trial for patients was NPN,
• 54:11and they received rocks.
• 54:12Luton Airport, just Jack.
• 54:14Start fires inhibitor over long periods of
• 54:17time is actually going opposite direction,
• 54:21so they found that these patients
• 54:23developed in 6% of nine patients out
• 54:26of 157 was NP ND well developed hybrid
• 54:30diesel lymphoma that were driven with Keras.
• 54:34And that's a 15 fold increase risk.
• 54:38So what they said in this study
• 54:41is that actually find one reason.
• 54:43Population of the Start 5 pathway enables.
• 54:47The transformation of the pre
• 54:49malignant B cell tumor that carries
• 54:51the Chaos Legion that essentially
• 54:52what we did in our genetic experiment.
• 54:55So I think it can cut both ways so it
• 54:58can be beneficial if he find ways to
• 55:00leverage this activity to completely
• 55:03suppress oncogenic signaling.
• 55:05But like Veronicas NPN study shows,
• 55:07it can also go in your opposite
• 55:09direction if you try to achieve
• 55:12long-term suppression of 1 pathway,
• 55:14you might inadvertently activate the other.
• 55:18So it's really interesting.
• 55:19'cause obviously if you're
• 55:20going to use ruxolitinib you,
• 55:22it's context may be very specific.
• 55:24I mean, I know it's a small proportion,
• 55:26but that's a pretty heart risk.
• 55:35Just waiting to see if any other questions.
• 55:39Well, I think we're you know,
• 55:40really, at the top of the hour,
• 55:42so you know, want to thank Mark is for
• 55:44it really is an extraordinary talk.
• 55:47That's creating so much insight
• 55:49into the biology of AB cells.
• 55:51Both respect to cancer and autoimmunity.
• 55:53Wanna thank Doctor, Snyder,
• 55:55and Snyder for sort of share
• 55:57continuing to lead this lectureship?
• 55:59And I want to thank the Frisbees
• 56:02for their continued support of
• 56:04our Cancer Center and the mission
• 56:06and the support of this lecture.
• 56:11I wish you all a great
• 56:13day and happy New year.
• 56:17Thank you. Thank
• 56:19you, thank you very much.
• 56:21Thank you Marcus. Take care.