Yale Pathology Grand Rounds: September 29, 2022

October 06, 2022

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Metabolic Impact on Tumor Immunity and Immunotherapy presented by Weiping Zou, MD, PhD

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00:06OK. We are on time 12:30,
00:08so we're going to start and
00:10so first welcome everybody.
00:12This is the first in person brand grounds
00:16meeting and we're very happy about it.
00:19So we expect to have people joining.
00:21We have 20 people online and maybe
00:24some more people to come so.
00:26So it's a great pleasure
00:28today to have our speaker,
00:30his doctor Weiping Zou.
00:32He's currently the Charles
00:34Denman Creek professor.
00:36He's a professor of immunology,
00:38pathology,
00:38biology and surgery at the University
00:40of Michigan and he's also the
00:41director of the Michigan Center of
00:43Excellence for Cancer Immunology,
00:45immunotherapy.
00:45And he has a number of additional
00:48appointments that I will omit now
00:50because I think they just add too much.
00:55Maybe he's training.
00:56He obtained his MD in China
00:59in Tongji University,
01:00and then he went on to get
01:03obtain a PhD in Paris.
01:04That was followed by postdoctoral training in
01:07France and then postural training in Baylor,
01:10Dallas.
01:12And then from his academic career,
01:15he initiated his career as an
01:18assistant professor in Tulane
01:20University in New Orleans,
01:21where he rose to the rank of
01:23associate professor with tenure.
01:24And then he moved to University of Michigan,
01:27where he became a full professor
01:29and director of the program.
01:32What is very interesting is that
01:35he's a very productive investigator.
01:37He has over 190 publications.
01:40I could count about 35.
01:42Really high impact publications in major,
01:46you know, sales, science,
01:47nature,
01:47type of journals and he's also very
01:50productive in the research and I
01:52could count 5R1 grants at this point,
01:54which as you can imagine is a huge
01:57amount of effort and shows to the
02:00reflects the quality of his work.
02:02He has done major contributions
02:03in the fields of tumor immunology
02:05and looking at different aspects.
02:07And recently he has focused more
02:09on the role of the metabolism
02:12and how metabolism can actually
02:14compromise adaptive immune responses
02:16in the tumor microenvironment.
02:18I have to also to say that he's
02:21a very translationally oriented,
02:22so his work focuses on very basic
02:25mechanisms but also projects into
02:28different tumor types and he has
02:30done really prominent contributions
02:32in colorectal cancer.
02:33Ovarian cancer and then also breast cancer.
02:36So it's a very, very diverse profile.
02:39And so without further ado,
02:41I believe Doctor Weiping Zou and his title
02:43is metabolic impact on tumor
02:45immunity and immunotherapy.
02:47And thank you very much.
02:59Right. First of all I would like to.
03:04Thank God for your kind
03:08invitation and introduction.
03:11Year is a pioneer institution
03:14of modern immunology,
03:16particularly human immunology,
03:19innate immunity and T cells.
03:24We paid up allergy,
03:25so in many ways this institution
03:28contributed enormously to our knowledge
03:32and also immunology translation.
03:35So this is absolutely a great
03:38pleasure for me to be here.
03:40This is the first time I'm able to give
03:43this talk to this prestigious university.
03:46As I mentioned yesterday when we had dinner,
03:50actually my old son wants to
03:52get into the C university.
03:55He failed and I emailed him.
03:59I said, I'm going to give a talk
04:01to this university and he said,
04:04OK, you succeeded.
04:06Thank you again for this
04:09wonderful opportunity.
04:10So.
04:12See you guys.
04:22So use too many minorities when we
04:26view our cancer therapy history.
04:30We have come a long way.
04:34First we have surgery,
04:36radiation, chemotherapy.
04:37Chemotherapy and the targeted
04:40therapy for managers and rotation if
04:42these days we do our immunotherapy.
04:45As you can see from this summary.
04:49Each milestone is really based on the
04:53basic celebs scientific discovery.
04:55So people always asked what is next.
05:01So in terms of immunotherapy,
05:05what we have done?
05:09Actually, early on we know based
05:13on the genetic identification and
05:16mutations people have discovered,
05:19it's pretty clear cancer
05:22is a genetic disease.
05:25But all work and many others work.
05:30Consider cancer is an immune disorder.
05:34And further we have studied the
05:37immunosuppressive mechanisms
05:38including the P1P1 that was in the
05:42human tumor migraine environment.
05:44So we believe the human tumor
05:48microenvironment holds the key to
05:50understanding human immunity and therapy.
05:53So at this stage,
05:54when we say these two contributions,
05:57conceptually speaking, it's easy.
06:00But when you talk about this 20 years ago.
06:04It's not the same thing.
06:07So we have some video articles
06:10in this space as we focus on
06:13immunosuppressive mechanisms.
06:19For those who are relatively new
06:22to terminology, as we know a lot
06:25of people getting into this area,
06:27you may see some of our work
06:29because we have not only reviewed
06:31the studies from our own group,
06:34but also from many others.
06:36As you may appreciate actually at least
06:392 high profile review articles we're
06:42done with Livingston I remarkable
06:46faculty are ADO Institution.
06:52Who's good mentioned?
06:54We are pretty much legislatively in the basic
06:58mechanisms as well as the transformation.
07:02Therefore. For all those years,
07:05we have been working on one concept.
07:08Say to again cancer microenvironment
07:10host key to understanding too
07:12many immunity and therapy.
07:14To address this we have several research
07:18directions or research angles you may say.
07:22For example you know
07:25suppressive mechanisms network.
07:27Such as PD1PD L one OK.
07:30Then cancer athletics and
07:33epigenetic reprogramming.
07:34And then a key immunologic pathways
07:37such as individual pathway,
07:39image C and stats.
07:41And finally,
07:42in the recent years we spent a lot of
07:46time working on metabolic pathways.
07:49So I guess I'm going to spend a little
07:52more time on the fourth direction.
07:55I will give you just one
07:58slight each for A&amp;B&amp;C,
08:00just to show you where I have come from.
08:08So and like to share this site with
08:10you for the first research direction
08:13we have immunosuppressive networks
08:15actually almost more than 20 years ago.
08:21Under the support and
08:23collaboration with Millington,
08:24we have published this
08:26paper in Nature magazine.
08:28It was named it is time PDL one.
08:31It's not me PDL one.
08:33It's named P 781.
08:35These people for the first time
08:38demonstrating PDL one of P-70 joint
08:40expression recognition and profit in
08:43the human cancer microenvironment
08:46and human human chain influence.
08:48We clearly demonstrated if you
08:51broke this pathway you can recover
08:54the dysfunctionality cells.
08:58This is far before the success of
09:02clinic trials, either with PD1PD1
09:05blockade or with anti serial four.
09:08Of course in these days if you look
09:11at the PD L1 you will not be able
09:14to find this table because early
09:16on when leaving discovered the best
09:19way he named this gene as B7H1.
09:22Of course, he has many other
09:24peaceful family members, as you know.
09:27So the second with your generation
09:29cancer at genetics,
09:30I know you have quite a few folks
09:33who are interested in epigenetic
09:35recognition in this institution,
09:38but we look at this from the
09:41immune recognition perspective.
09:42So in the tumor microenvironment similar
09:45to the TH1 and TH2 reciprocal recognition?
09:50We have observed a reciprocal
09:54regulation between PRC two complex
09:56and Swiss sniper complex in the tumor.
10:00So actually this recognition was
10:03properly controlled interference
10:05zeronine therefore TH one type second
10:08production and T cell trafficking
10:10and human energy.
10:11We have worked out the detailed
10:15biochemical and functional mechanisms.
10:17We I'm not going to show you
10:19the details as I mentioned.
10:21Then third, research direction,
10:24I know also several investigators
10:27including my host code is very
10:30interested in this image C interferon
10:33signaling sets signaling pathway.
10:36As you know these are the key immunogenic
10:39pathway in the immune responses,
10:40not only just in the tumor
10:42migraine environment.
10:43I show you one example.
10:44We have,
10:45we have discovered actually you know
10:48mutations in MHC pathway and stat
10:52and interferon pathway are considered
10:54a celebrity resistant mechanism.
10:58But we know the vast majority of the
11:00patients do not have those mutations.
11:03Therefore we need to find out
11:05the other pathways which may
11:08contribute to safety resistance.
11:10So one pathway we have discovered.
11:14Actually,
11:14the integrity of interferon
11:17signaling pathway is controlled by.
11:22Jean quota of January.
11:23So this is a auto between septor.
11:27It turns out actually all of the nearing
11:30can control the stability and the
11:33degradation of the film gamma receptor.
11:36So as a consequence,
11:37this controls the image C expression antigen
11:40presentation and the T cell functionality.
11:42OK, I don't have time to
11:45show you this was published.
11:47For those who are interested in this,
11:49you may have a look instead.
11:51Most of my time we are focused on the force
11:55we switch direction metabolic pathways.
11:59And I will.
12:02Talk about the basically two stories.
12:05One is system XC and CS4,
12:09its relationship with tumor
12:12cell philippoussis.
12:13Another is SLC 43A2.
12:17So those ACC family members are
12:20nutrients or metabolite transporters.
12:23There are several hundreds of them.
12:25Most of them are poorly understood
12:28in the field of immunology.
12:30We start to figure out some of it.
12:35So before that I want to introduce
12:39the concept of ferroptosis.
12:41So it has been defined in vitro
12:44through the synthetic compounds.
12:47It means the cells will die
12:50through iron dependent but lipid
12:53peroxidation induced cell death.
12:56There are several genes or pathways
13:00associated or regulated cell biosis.
13:03So,
13:04including this exit system
13:07GX4 and a CSR four.
13:10However there is no CBC marker
13:14to define Philippoussis.
13:15What we usually do we use a few
13:19criteria to define electrolysis,
13:21so one it's maybe the Rose production,
13:24another is expression of all states
13:27needed species on the membrane,
13:29and finally we need to see
13:32the functional activities.
13:34So in this case we asked
13:37a very simple question.
13:39We know when CDA T cells are activated,
13:42engage tumor cells.
13:45CHT cells we need preparing,
13:48makes pores on the membrane,
13:50then grant them.
13:51We get into the cells activated
13:54caspase induce tumor cell able doses.
13:58This is text book message.
14:02So we asked a simple question means
14:05if CDA T cells who kills tumor cells?
14:09This is a way how the tumor cells
14:11die is Philippoussis involved.
14:13So in this case we set up several
14:17experiments to test this possibility.
14:20So one is a 88 over retention model.
14:24We do immunotherapy before you can see PD,
14:27one can control the tumor growth.
14:31Under this condition before the tumor cells,
14:34really it is an 8 stage before
14:36the tumor cells die,
14:37you give the tumor cells out,
14:39you detect relative needs worse production.
14:43You see actually the immunotherapy
14:46induces liberals production in
14:48the tumor cells in PD1 cell.
14:51Then we did a T cell therapy model.
14:54Be 16 over expression cells and
14:58treated in vivo with only one cells.
15:01It's not a surprise tumor is controlled
15:04and again we see individuals production.
15:06So this suggests maybe T cells or immune
15:10therapy can promote liberals production.
15:14Maybe in May induce Philippoussis
15:17tumor cell paralysis.
15:19We did in vitro studies in this case
15:22to provide direct evidence we cultured.
15:2541 cells with All Blacks fashion tumor
15:28cells we look it's rose production
15:30again we see rose production induced
15:33by by only one cells and this antigen
15:36specific away can see more but if
15:39you look at the tumor cell death.
15:43Of course you use a small amount
15:45of T cells and in this case you can
15:48have space to manipulate the system.
15:50You will see you your small amount
15:52of T cells. You see T cell cleaning.
15:56And under this condition,
15:57if you use a small amount of RS3
16:01is a dosis inducer,
16:03you'll see some levels of human cell death.
16:06If you put them together,
16:07you see dramatic human cell death.
16:09These tumor cells can be
16:13completely abolished.
16:14By THEODOSIS inhibitor,
16:17So what we have here, it means T
16:22cells can promote tumor cell factories.
16:26However,
16:26T cells themselves are not sufficient.
16:29You need to have a trigger somewhere.
16:32So we will explore further about
16:35this phenomenon.
16:36But we studied further in vivo
16:39in military condition.
16:40So it's a classic model.
16:43For example, with P-16,
16:44if you treat the tumor bearing
16:47mice with antiscia 4 and PD1,
16:49you'll see very nice tumor control.
16:52But if you treat the mice under
16:54this condition with liberal statin,
16:56it's a Philippoussis inhibitor.
16:59The therapeutic efficacy
17:01is basically punished.
17:02So this is very unusual.
17:04This is completely unexpected.
17:07Because we all know
17:10CD8T cells kill the tumor
17:11cells through able process.
17:13How come a ferroptosis
17:16inhibitor can Polish the effect?
17:19To really ensure this possibility,
17:22we used another model means we in vitro
17:26generate erskin resistant tumor cells.
17:29It's similar to chemotherapy resistant cells.
17:33You do the individual generates the
17:35cells with this to the reverse inducer.
17:38Then you do the immunotherapy.
17:41OK you see here,
17:43parental cells are responsive
17:45and resistant cells are no longer
17:48responsive to immunotherapy.
17:50Indicating actually Ferroptosis is a
17:55potential mechanism induced by immunotherapy.
18:00So we look it's Morgan mechanisms
18:02then to make a non story short,
18:05we know it's interfering and other things,
18:07but just show you interfering here.
18:09If you make a lookout receptor in
18:12the knockout tumor cells and your
18:15culture with only one cells you will
18:18see actually the liberals production
18:20in the human cells is basically gone.
18:23Under the tumor cell death is
18:26also basically gone,
18:27so indicating this tumor cell death
18:30is controlled through the interference
18:32and interference signaling.
18:34So then we hypothesized maybe
18:38interfering with your stimulate
18:40oxygen lipid species and therefore the
18:43cells die as we previously mentioned.
18:47So we did some individual studies.
18:51We cultured tumor cells with our CS
18:54refill processing user with or without
18:57anything comma you can appreciate.
18:59In fact the aim is without
19:03show actually interfering.
19:05Gamma can induce tumor cell
19:08oxides lipid species,
19:10so particularly USB 16P18 or induced.
19:13So this is increased when
19:15you have a small amount of.
19:17ISL 3.
19:19So it means actually in the film
19:23gamma can properly do the job.
19:25To directly show this we used a independent
19:29comma sensitive human tumor sauna.
19:31It's HD human cell line
19:33you treat with interferon.
19:34You can inhibit tumor growth
19:36in the English model,
19:38but under this condition if you use
19:40liberal studying you will see the effect.
19:42It's completely gone.
19:44But Earth asset in vitro if you only have
19:47interference or you only have 3 cells.
19:49The sale gas we are not having,
19:51so suggesting something else,
19:53not only just in.
19:55So but what did anything do in this case?
19:59We look at the molecular targets,
20:01potential molecular targets of
20:03interferon particularly XC system.
20:05As you know XC system can transport
20:09system into the cells then become
20:13system and GSH and this will protect
20:16the cells test from the tosis.
20:19So it turns out interferon actually we press,
20:24we press the exit. System.
20:26So this is just the the among a.
20:28This shows you the protein.
20:30Not only this,
20:32it's functionally important as shown here,
20:35because the system update is reduced
20:38when you have independent treatment
20:40and then the GSH synthesis is reduced,
20:44and particularly if you have
20:46again small amount of erosion.
20:48This is a well known reduces GSH
20:50when you put them together though
20:53reduction of GH is really dramatic.
20:56So we extend our studies to humans,
21:01not only just we use the human cells,
21:03human human cells and we meet a
21:06correlation with immunotherapy.
21:08As you can see here when the
21:11patient received
21:12e-mail service called Panini Benefits,
21:14the XC expression is done in the
21:17tumor of course, the interferon
21:19signaling and CTA is increased.
21:22So what do we have?
21:24At least we can say apart from apoptosis.
21:29The interaction between CHT cells and
21:32tumor cells in this context fail.
21:36Photos may be involved,
21:38and interferon gamma can target
21:40excision to be involved in this space.
21:43This has not been previously
21:45appreciated because we don't know.
21:47We only think that this is able to process.
21:50So now as I mentioned in the film,
21:53comma is not enough,
21:54T cells are not sufficient.
21:56So what else?
21:57What else?
21:58Because early on when Phil poses
22:01as a concept was was established,
22:05it is basically based on the
22:08synthetic compounds.
22:09So you treated the cells with the chemicals
22:12and then you see the philippoussis,
22:14you see the pathway.
22:16So if their process is a intrinsic mechanism.
22:20We should have a intrinsic
22:22mechanism to induce the fibrosis
22:24in the cells because we don't have
22:27synthetic compound in our body.
22:28So we look for the natural
22:31theodosis inducers in.
22:33So in this case we come to a fatty acid diet.
22:38So the reason is we know they are
22:40quite many publications talking about
22:42the relationship between bias and the
22:45celebrity response to immunotherapy.
22:47They are also quite some
22:50publications talking about.
22:51Micro got microbiota and tumor cell
22:55respond where patient responsive
22:57responsiveness to immunotherapy.
23:00So therefore we were thinking
23:02maybe interfering is one thing,
23:04maybe some my tablets some
23:08metabolic nutrient will be involved.
23:10We turned to fatty acids because
23:13we know when the cells die through
23:15their process,
23:16it's because of oxidized lipid species.
23:18That's why we look at the fatty acid.
23:21So then I invite you to look at
23:23several groups of fatty acids.
23:25So in fact you have short chain,
23:27medium chain,
23:28non chain and a very long chain fatty acids.
23:31I want you to pay attention on
23:34the non gene fatty acids such
23:36as POAOA and arachidonic acid.
23:38Here we checked all of it.
23:40So in this case when we look at the
23:43map of fibrosis people have defined as
23:47a fibrosis involved genes and one is called.
23:51A CSR 4 here.
23:52So in fact that you prefer it's
23:55an enzyme preferred substrates,
23:57it's electronic acid AA.
23:59So finally you will see the final product
24:03is Poly unsaturated offset lipid species.
24:06So in this case we are
24:08thinking it should be involved.
24:11So what we did,
24:12we cultured the tumor cells with interferon
24:15pronounced different fatty acids.
24:18Long term,
24:18short term media change often.
24:20Then we look at the cell desk.
24:22It turns out that in the presence
24:24of a the tumor cell,
24:26death is dramatically increased.
24:28And keep in mind that these
24:31cell deaths can be completely
24:34blocked by THEODOSIS inhibitor.
24:36So it means this is really theodosis
24:38and this is repeated reproducible
24:40in P-16 and seven tumor cells
24:43in mouse and humans.
24:45So finally we want to see what has
24:49happened actually with electronic
24:51acid in the presence of in the
24:54field. So we cultured human cells
24:57with interfering with or without.
25:00E5 neighbored atonic acid we want
25:03to see where electronic acid goes.
25:07So in this case we made a CSL
25:10knockout and the width of tumor cells.
25:13We treat the cells in this way.
25:15Then we look at different oxygenated
25:18species because you may appreciate
25:20here what is the black box and
25:22the red bars are all deficient.
25:25It is deficient cells.
25:27You will see actually interfering,
25:29really promote.
25:31The incorporation of T5
25:34neighboured electronic acid in two
25:37different oxides lipid species.
25:39So this including PE18B16 PC 18.
25:44You can see from the slight ACL 1400.
25:48So in this case,
25:49what has in the gamma Dong look at the
25:53brooding expression of CCL 4 actually
25:56in film comma stimulate its expression,
25:59so this is slow transcriptional
26:02recognition as the cheap essay shows.
26:05Actually there's a high I funding
26:08in the ACL 4 promoter area,
26:11and the cheap shows actually
26:14this high occupancy.
26:15So we first did some in vivo
26:18studies to show the relevance.
26:20So in this case we made a ACL 4 knockout
26:24tumors in several tumor cell lines.
26:26You see a CS4 is gone and tumors are
26:29getting bigger and in vivo and when
26:32we did the combination therapy Ravi,
26:35Classic way we treated mice with AA and
26:38AA actually can partially control the
26:41tumor progression in several models,
26:44but keep in mind the A.
26:46He's a very small amount of concentration.
26:48You you cannot give too much and then
26:50you kill the mice because it's quite toxic.
26:52So we see the combination therapy can give
26:55you some benefits in the mouse model.
26:58So when you look at the patient
27:01with a CCL 4 expression,
27:04in fact high ACC for expression is
27:08positively associated with patient survival,
27:11suggesting maybe it is a four is relevant in.
27:16The tumor microenvironment.
27:17So we tested it not only like in tonic acid,
27:22we tested them other fatty acids
27:24as I mentioned,
27:26but what I conclude here don't
27:28show you all the details.
27:30Apart from AA,
27:31OA and POA can also participate in
27:36inducing the tumor cell process.
27:40All these essays are in the absence
27:44of synthetic compound.
27:45So indicating what we discovered actually.
27:49The effect specific fatty acids plus
27:52interferon gamma are the intrinsic
27:55philippoussis inducing mechanisms
27:57we are able to detect.
28:00Of course all the fatty acids
28:02species and interference in vivo,
28:05they are not synthetic combo.
28:07So this is another similar to
28:09the concept that we all know,
28:12such as H170 cells, not one cytokine.
28:16It's not enough to polarize,
28:17you need several.
28:19Effectors what we have discovered, actually.
28:22Tumor cell Philippoussis needs
28:25several factors,
28:26and interference is one of them
28:28and the fatty acids are another.
28:31So now that's the the the conclusion
28:36we have basically when you have the
28:40induction between C8 and tumor cells.
28:43Because this is one of the founding
28:46father of Tosis is another.
28:48I hope this becomes textbook.
28:50So fear of loss is is mediated
28:53and the recognition
28:54through the AC system and CSL 4.
28:57Maybe other factors will be involved as well
29:00and we are still working on the details.
29:04As you know, there are several
29:08philanthropic pathways people have defined.
29:12So what is the technical message here?
29:16You must fear of those is is
29:19a mode of action of Syria.
29:21And tumor Philippoussis is
29:24neural therapy mechanism.
29:25So if so we should think about
29:29the potential translation.
29:31We are thinking about this,
29:33many groups are working on this.
29:35So now we move to the second part of my talk.
29:38It's concerned another ACC family member,
29:42it's named SRC 4382.
29:45So myself is a immunologist and when you
29:49talk to biologists and some other people,
29:54there is an idea or thought proposed
29:58because the tumor cells are highly
30:01proliferative and invasive.
30:03The tumor cells need a lot of nutrients.
30:05So one way to treat the patient
30:07that maybe we can start with the
30:10cancer cell death.
30:11So that's the way how the
30:14biology is maybe some.
30:16Pharmacologists think this way,
30:17I don't know, so let's see if this works.
30:21In that case,
30:21I invite you to think about the
30:23nutrients and metabolites in
30:25the cancer microenvironment.
30:27We know when the cells are exposed to
30:30different metabolites and nutrients
30:32in the particular environment,
30:34not only just human cells,
30:36but also these cells and disease
30:39and Macy's and other cells,
30:40they must be subject to the
30:43regulation by the environment.
30:44Therefore, they are functional.
30:45Status must be changed.
30:47So it's a very simple way to
30:50put so in this case.
30:52Early on,
30:53some groups have already discovered
30:55the T cells are dysfunctional
30:57in the tumor micro environment.
30:59You may say the T cells are exhausted.
31:02That's alright so.
31:03But we also know some epigenetic
31:05pathways are involved in the regulation
31:08of tumor cell dysfunction and T cell
31:11dysfunctionality in the tumor environment.
31:14So we are thinking maybe in this
31:16case a crosstalk between metabolic
31:19and apologetic mechanism.
31:21This has, uh, evidence actually.
31:24People have reported some of them.
31:26For example,
31:27you know,
31:27after H succinate and have a
31:29particularly succinate have has
31:31been studied in macrophages,
31:34minor cells and some others.
31:36And we are interested in Sam.
31:38So why we are interested in them,
31:40you will see why we're interested in them.
31:43So in fact,
31:44in this case we look at the amino acids.
31:48So we did a very simple array,
31:52so we cultured basically G cells with
31:57different amino acids in the media,
31:59but we manipulated the concentration
32:01but we admit one by one and then we
32:04take the functionality of the T cells,
32:06basically the shell gas and the T cells.
32:09It turns out if you own it,
32:14my theory.
32:15So T cells cannot stand,
32:17they become very much evolution.
32:20And the cells do not express much
32:24interference.
32:25And then we did another way along
32:27means we add amino acids back.
32:29So it's a plus experiment.
32:32So in this case we calculate
32:34the cells with too much
32:35to induce the cell or
32:38dysfunctionality dysfunctional.
32:39They become embodied and reduce stereogram.
32:42You will see under this condition
32:45if we add methionine pack we
32:48will see actually the tumor.
32:51T cell F is reduced,
32:53T cell function is improved.
32:56So indicating actually the T cells
33:00are very sensitive to the supply
33:04of methionine so and then we look
33:07at the methionine metabolic cycle.
33:09So in fact my theory can become same
33:13and you know Sam is a real donor for
33:19methylation so history modification.
33:21So that's the reason we want to
33:23look at some right so in this
33:25case when you cut your.
33:27Have to keep cells with two
33:29measurement and you supplement
33:31with my film and you detect all the
33:34Internet intercellular my cabinets
33:36you will see if you do so first of
33:39all you see reduced intracellular
33:41methionine when you don't add in
33:44the culture and when you add it
33:47comes back and also you don't have
33:50them in the SH all those things
33:52but you supplement Matheny you can
33:55partially and important we cover.
33:57They are the intercellular ascend
34:00and intercellular other metabolites
34:02of of methionine, such as SH.
34:06So if so this must affect his notification.
34:10So when we in this case we look at this,
34:13this is calculated with either tumor
34:16cells mouse are not human you will see
34:19actually the tumors supernatant reduce
34:22its 3K790 resonation dramatically.
34:24This is not only the case in in most cells,
34:27in too many cells the same thing
34:29and you supplement methionine
34:31which you can recover it.
34:32Other histone markers are less affected and.
34:36I I couldn't explain to you why.
34:38Then we look at the primary cells T
34:41cells in the tumor micro in humans
34:44and the mouse and you will see
34:47isolated cells from the mouse system.
34:49You will see we used H3790 resonation
34:52and so is in human CD8T cells
34:57in the tumor microenvironment.
34:59So in that case,
35:01this extreme case 7 and T machination
35:03must be functionally important.
35:06So to test this possibility,
35:08we made a total of 1 specific
35:10lookout in T cells.
35:12The reason is total one is the
35:15only endemic 8719 resolution.
35:16So when we made it look out in
35:19T cells and then the tumors are
35:21getting bigger than the T cells are
35:24becoming apoptotic and dysfunctional.
35:25So that's one way to go.
35:28Another way to go is.
35:29We supplement methionine in the
35:31tumor bearing mice in this condition.
35:34If you supplement then you reduce
35:36tumor growth,
35:37you will cover histone modification
35:40in T cells and also you recover
35:43the T cell functionality.
35:46So we did it not only in mouse model,
35:48we did in patient with cancer.
35:50So we supplemented methionine to the patient.
35:53Then we see the T cells.
35:55It turns out if you do so.
35:58My third supplementation can we
36:02cover each 3K79 demethylation
36:04we cover even Step 5 expression.
36:08We checked all the other stuff
36:09because step five is most obvious.
36:11And furthermore,
36:12if you look at the second expression such
36:15as IO2, so before therapy, after,
36:18before, and after, you will see I2
36:21is largely recovered in T cells.
36:24You know, somehow step five really
36:27controls the expression of.
36:29I of O2, then we first look
36:32at the possible mechanisms.
36:34So it turns out actually it's
36:38379 emanation target Step 5,
36:40particularly step 5B,
36:41and the cheaper essay shows this is the case.
36:44In fact, if you cut yourself
36:46with supernatant and with
36:48the maternal supplementation,
36:49methionine supplementation can recover
36:52the occupancy in the certified model.
36:56So this just show you again,
37:00not only we cover the T cell,
37:03the the the,
37:05the the the the cheaper and also show
37:09H3K79 nations we covered and instead
37:11of having is recovered and I2 is
37:14recovered in both humans and mice.
37:17And finally we want to understand.
37:20If methionine is there,
37:22why the T cells cannot get better?
37:27So maybe the tumor cells all
37:29compete T cells for methionine
37:31in the tumor microenvironment.
37:34We turned our attention to methionine
37:38transporters so we screened all of them.
37:41It turns out actually you will see
37:45compare tumor cells and T cells in
37:48the same environment and actually
37:50the tumor cells express quite a lot
37:53of ACC for 3A2 is one of methionine.
37:57This product,
37:57so this is among A and this is protein.
38:01So this is T cells and many
38:03other transporters are similar,
38:05but they are quite some differences.
38:07So we are we continue to work on
38:10this space to define the different
38:12differences we are able to see and
38:15then to see the functionality.
38:17So this suggests maybe
38:21AC43A2 easy transporter highly
38:23expressed in the tumor cells is
38:25functionally important if so.
38:27We make a knocking down SLC 43A2.
38:32In the commercials then we start to
38:35capture the human cells with cells.
38:36OK, so you can see actually the T
38:39cells are becoming less able to
38:42reach the functions are recovered.
38:44So indicating ACC for this 382 is important.
38:49To further demonstrate this possibility,
38:51we did in DEVO studies,
38:53if you shut down PC police
38:55382 in the tumor cells.
38:57USC actually the tumor is smaller
39:00in the immune competence system.
39:03The key cells in terms of their
39:05number and their function are better.
39:07This is not only in one model.
39:09In several models we can see the case.
39:11So what we have here is a summary we
39:15see in the tumor microenvironment,
39:18tumor cells express high levels
39:20of transporter.
39:22For methionine it's ACC 4382
39:25outcompete T cells 4.
39:27The only surprise when T cells do not
39:30get methionine and the T cells have
39:33insufficient sense Earth myself honor.
39:36Therefore they cannot successfully
39:39do the H3K790 maceration and
39:42therefore regulate stats fab.
39:44And as a consequence this affect the TCL
39:47functionality and the T cell survival.
39:49So what we suggest here maybe
39:51you know we can either we do
39:54mathematics supplementation, we do,
39:56we cover the T cell immunity.
39:58Maybe we can particularly target
40:00the tumor as you C for 3/8 to
40:03the rescue T cell functionality.
40:05So now it comes back to the question
40:09we asked. So can we stop themselves?
40:13Can we stop to myself, to this?
40:15Yes, we can.
40:16You must ask really needed method.
40:18Earth one example.
40:20But the poverty is if you stop yourself
40:23to death, you also stop T cells to death.
40:26Under the AIDS and who kills the tumor cells?
40:30The T cells?
40:31Tumor cells.
40:32So that's why what we say.
40:33If you want to stop human cells
40:36to test using this approach,
40:38probably you kill 1000 yourself defeat
40:41800 and I would put the opposite way,
40:44you kill 800 yourself defeat defeat 101,000.
40:50So that's why we need to be really
40:53smart to consider not only just.
40:56To, to the tumors,
40:58but we have also considered the T cells.
41:01So now we ask the question again
41:03as I put it at the beginning.
41:05So what is next in terms of telling what
41:09is the next generation of cancer therapy?
41:12So in my view,
41:15immune therapy we means the basis.
41:19Why? Two reasons.
41:20Because the military has been
41:23successful to cure some patients.
41:25We already know this,
41:27indicating the powerful reason.
41:30The whole powerful is immune
41:32system could be second.
41:34T cells. Can kill human cells.
41:37It's not a surprise.
41:39And the further T cells remember to kill
41:42tumor cells and then nobody else can do that.
41:45So that's why I feel the next
41:47generation of cancer therapy,
41:49immune therapy, is the basis.
41:52So I stop here.
41:54I appreciate the contribution from several
41:57very tentative federal as I did here.
42:00Some of them are faculty,
42:02some of them moved to different institutions.
42:05And of course I didn't particularly
42:08talk about the PD1 video one study and
42:11some others it was a collaboration
42:13with and I have quite a few,
42:16some few other collaborators
42:20in the United States and.
42:22And in other places,
42:23thank you for your attention and looking
42:25forward to your comments and questions.
42:35Question.
42:38Thank you over here.
42:40So thank you very much.
42:42Much appreciated.
42:43Presentation understood correctly,
42:45you showed that the rock, correct?
42:47Doc acid was a mediator of T cell.
42:57You are. Has anyone looked
42:59inside the cell, two per cell,
43:01that's undergoing the fructose this
43:04kind of objectively by the elements for
43:06others to see what's elevated threat?
43:11It is surveyed, but there
43:14are thousands of. Liberal.
43:17Right. So in response,
43:19So what we have done actually we
43:23detected electronic acids in the tumor
43:27microenvironment in the tumor floats.
43:31So the question is very tricky.
43:34You have to have sufficient
43:36levels of electronic acids,
43:37but if you have very high concentration
43:40you kill everything. Right.
43:41So, so, so The thing is you have
43:44to have two things simultaneously,
43:48one is interfering, another is doing acid.
43:52So that's a play you have to go.
43:55Yeah, we did not systemically to
43:58detect all the metabolites in the
44:01tumor microenvironment by our own.
44:03There are some report in that space.
44:05There are technical challenges
44:07in in that situation.
44:09I guess the question is what do you ask?
44:11It's a it's very annoying to to do it.
44:13For example,
44:15we really dynamically monitor the
44:18metabolic environment when the sales
44:21either become able to or philanthropic
44:25whether there's any difference.
44:27In this case, we need to have props.
44:31It's a real property.
44:33We follow these people.
44:34Maybe some of you are smart and and
44:37have tools we can we can do that.
44:39So one day it could be done.
44:41You know, just a trace where it
44:44goes and how high the levels are,
44:48yeah.
44:49Thanks.
44:54So I wonder if the opposite guys.
45:04Struck.
45:16Great question actually.
45:18So as far as my understanding is,
45:22you know when we look at the Philippoussis.
45:27It's all the different ways how
45:29the cells die or four for example.
45:31You know you you have some
45:34executive genes which have been
45:36well defined in enable those poses,
45:38right but for fear of nosis,
45:42it's really about the membrane and
45:44damage so mediated by oxygenic
45:47species that's what we know.
45:49So therefore so very direct question when you
45:53asked whether the factors what we studied.
45:57Or whoever studied have
45:59directly effect on the membrane,
46:01let's say maybe the nails and the structures,
46:04those things, right?
46:05We didn't go that far.
46:06I even don't have the expertise to do that.
46:09So, so I think it's very nice way to go.
46:13So one way to to do it is we have
46:15done a little bit means we detect
46:18a tumor membrane if it oxidizes,
46:20inhibit species, that's what we know.
46:21But we don't look at the structure
46:24to monitor how the sector changes
46:27could be unacceptable.
46:28Yeah, but we didn't know that.
46:30We even don't know how to do that.
46:32So maybe.
46:34Which way we go that maybe again,
46:36if we have some proxy,
46:38it could be useful, right.
46:40So maybe you, you,
46:41you have some ideas in that space.
46:42We were chatting it on maybe this,
46:44that that's a good way to go
46:46because if people are still some
46:48people feel or feel to see if
46:51you don't have a executive gene.
46:53So what are you talking about, right.
46:54But The thing is the pathway is regulated
46:57and the pathway can be inhibited,
46:59can be activated, it can be regulated.
47:01So that's very difficult mechanism, right?
47:03So therefore it's.
47:04The program still this.
47:22Yes, yes, yes.
47:23That's also a good point.
47:25So you know, when you design experiments,
47:28you want to see something,
47:29you look at something, right?
47:30So therefore we didn't look
47:33at some other cells.
47:35So my. So right now we know
47:38different types of cells.
47:40We have given the sensitivities
47:42to different stimuli,
47:44stimuli, fibrosis stimuli.
47:46They may have different
47:48mechanisms to control.
47:50So that's typically something we are
47:53working on including for example,
47:55how about megabytes?
47:56How about T cells, right?
47:58So I guess this is not an mechanism
48:01exclusively for tumor cells.
48:03There is no such thing.
48:05So the mechanism could be functional
48:08for other types of cells.
48:10The question is?
48:11When and how and which one?
48:14We are working with this
48:15but you can go on details.
48:20This idea the only competition
48:22out of competition
48:23of tumor cells.
48:26Do you think it's the growth across
48:29towards you know we think about like
48:31lung which is sensitive to prepare
48:33incorrectly that is not or for example
48:35the location of the tool you have
48:37the two reason delivered reason the
48:39London or different access to metabolic
48:41substrate have you what are your thoughts
48:45about that? Yes it's it's OK.
48:46It's a great question.
48:47It's hard to address. OK.
48:49So what we started to look
48:51at since even in the same we
48:53have recently paper just.
48:55Eventually you even in the same
48:57human parent such as liver right?
49:00We reach locate the neighbor metastasis HCC.
49:03So in the liver microenvironment
49:06you have HC and metastasis.
49:09Then we look at even the same
49:12macrophage subsets they are
49:14metabolic patterns are different.
49:17So if they are, metabolic patterns
49:20are different, therefore their
49:22metabolic needs are different.
49:24How does it happen? Right.
49:27We have nuclear. You know,
49:29but we are still working on those things.
49:31So we we work more on the the way how the
49:36sales die because we believe this is this,
49:40these matters are not.
49:42So so the thought is.
49:46Different cells have
49:48different metabolic pattern.
49:50The same cells in different metabolic
49:53environment may have to adapt this
49:56particular environmental survive #1, right?
49:58So then whether they can expand.
50:00So I guess a part from the genetic
50:05mutations which people have started
50:07or not in the space of cancer biology
50:09and genetics in the recent days,
50:11people really moved to the
50:13field of metabolism because the
50:15metabolism is somehow universal.
50:17It must be regulated in
50:19one way or versus another.
50:21So that's that's why I we we have
50:23high interest in this, right.
50:25But the answer is very big.
50:27I know I didn't really give you an answer,
50:28just what we have.
50:33Specificity. Signing on.
50:38So why do you think?
50:43Yes, that's, uh,
50:44it's it's a great question actually.
50:46It's. So it turns out this is a, it's a,
50:50it's a, it's a biochemical question.
50:53OK. So if you look at the constant,
50:57it's the lowest among all the
51:01other isomorphic modifiers.
51:02So that's why it's. So it's sensitive.
51:05It's. Yeah, yeah, yeah. Yeah.
51:08Yeah. So so this actually this.
51:11This information is available.
51:12It's not from us,
51:14it's from when we figured out that it's
51:17needed on and we asked the same question,
51:19ask to ourselves why we
51:21see this is predominant,
51:22the others are not so dramatic and
51:24then we know it's publications.
51:26It turns out that's the case.
51:27Yes.
51:31In any of your models you're looking at.
51:35The cancer cells undergoing sister type.
51:41Um, I working on this.
51:44We are working on this.
51:46We should though I'm
51:48not, I'm not working on the persistent.
51:50So, so yes, it's a great question.
51:52Actually we initially I was really puzzled.
51:56Puzzled by what when you see I show you
51:59the picture actually when you treat
52:01the mice with PDL one and the CDA 4.
52:06And under this condition you treated
52:09mice with fair process inhibitor?
52:12And as the therapeutic efficacy is gone.
52:15This puzzled me so because, I mean,
52:18we we we know this is able to see
52:19the T cells kill tumor cells.
52:21It's even though this is caspase,
52:23and it's very well established,
52:25you cannot throw away all those
52:26things what people have known, right?
52:28So the only explanation is these across.
52:33So who is first, who is second,
52:35who is in the middle and who initiate what?
52:37Who emphasis what? Those kind of things.
52:39So we worked very hard,
52:41but we have no group so far.
52:43But we know there must be a gross.
52:45Yeah, I'm thinking in terms of like the
52:47paper from the green script where, you know,
52:49they show that the many monitors slight
52:56differences.
52:59Yeah, that's a possibility. Uh, actually,
53:01I just had a discussion recently.
53:04I probably will discuss with him
53:06again to see which way we can we
53:10can get some insight. Yeah, yeah.
53:14So I have another question
53:15about practical you know,
53:16we ask pathology for always
53:18frustrated by PO1 as a biomarker or
53:21TMB like there's no good biomarkers.
53:25It sounds like you have about a number
53:27of potential molecules that could work
53:29as biomarkers, you know having the
53:31right transporters in the right place.
53:34Do you see any, any sort of immediate
53:37possibility of some of these as
53:39biomarkers for immunotherapy? Yeah, so.
53:44So I I guess this is is it's quite a
53:48it's quite a depressing I would say.
53:50So when you located the biomarkers right,
53:53so it could be money you know perfectly
53:56well when people started to do the PD1
53:59PDL 1 clinical trials and nobody knows
54:01it the PD one or PDL one expression.
54:04So now after that you know it is PDL
54:06one expression and if they approve
54:08you know you have certain levels of
54:11PD1 expression it's indication right.
54:13So it's. It's not the way how how we know
54:17initially for it is for Philippoussis.
54:20I don't know which one we can
54:22we can really say.
54:23So the best way is well check all the
54:27social associated genes particularly protein
54:30levels whether this will give us something.
54:34You know so for example we looked at
54:38a CSR four expression when you see
54:41high ACR four expression may this may
54:46help can it is for BA real bellmaker.
54:51You get to have something to test it
54:53in clinic, in patient and your mouse,
54:55you know mechanism, fine.
54:58But you if you want to see it directly
55:00in patient, that's another story.
55:02We need to see the patient.
55:04That's why we appreciate your work.
55:06We need to see the patient.
55:07We need to see the tumors in
55:09patient and see what's going on.
55:11Yes, but there are ways to go.
55:57OK.
56:08I'm I I'm afraid I really didn't get
56:10any question you asked. Maybe just.
56:40So you mean when they express the grammar?
56:45Season.
56:51Right.
56:54So that's why. Our fear of dosis,
56:57it's obvious when we do immunotherapy.
57:01So that's the system we used.
57:03So actually in response to
57:05your question early on when you
57:08just give tomatoes and mice.
57:10So both you still have T cells and T
57:13cells are more or less functional.
57:15But under this condition we treated
57:17myself with ferocity and crypto.
57:20We hope, we hope we can see.
57:23We thought.
57:25So you get to have sufficient levels of
57:29interference and electronic acid and so
57:32and maybe other BOA&amp;OA in the environment.
57:35How do we do that if you don't have
57:40a sufficient T cell infiltration?
57:43Even so, I guess we can manipulate
57:47the system, for example,
57:48maybe for example you have some,
57:50it's not one way to go, maybe for example,
57:52if you have a card in cell therapy, right?
57:54So not all the patients are responsive
57:57and then we have some cells here,
57:59maybe we can manipulate this
58:01specially in this, in this way,
58:03another way we can do also maybe you know we
58:06have ways to improve the teacher trafficking,
58:09right?
58:10So, so, so far where do?
58:15A pure airplus mechanism.
58:17In the absence of immune system
58:19whether this is a valid approach, we don't.
58:22Maybe there's a way to go?
58:26Maybe radiation or chemo or something?
58:29Yes and a no.
58:31And also we have another paper I can
58:33mention this here and we have a paper
58:35to actually that's the first people
58:38talking about the effect of radiation
58:41is partially dependent on our fibrosis,
58:45but this fabulous especially again
58:47recognized by the immune system.
58:50So you need to have an immune system.
58:53Yes,
58:53so that that's a cancer discovery people.
58:57We.
58:57Properties 4-5 years ago,
59:00yeah.
59:02Questions?
59:06Thank you very much
59:06again. Yeah. Thank you.
59:07Thank you. Thank you.