Pathology Grand Rounds, Sept. 25, 2025 - Peng Ji, MD, PhD

Transcript

• 00:01It's my great honor to
• 00:02introduce Doctor. Peng Jie as
• 00:04our Grand Rounds speaker today.
• 00:07Doctor. Jie is a physician
• 00:08scientist,
• 00:09and tenured professor of pathology
• 00:11at Northwestern University,
• 00:12where he serves as vice
• 00:14chair for research and holds
• 00:15the Marie a Fleming
• 00:17professorship.
• 00:19His work is advancing experimental
• 00:21hematology as well as clinical
• 00:23hematopathology.
• 00:24After earning an MD from
• 00:26Peking University,
• 00:27PhD from Albert Einstein, and
• 00:29completing a postdoc at MIT,
• 00:31he joined Northwestern's pathology physician
• 00:34scientist training program in twenty
• 00:35eleven, later gaining board certification
• 00:38in clinical hematop clinical pathology
• 00:40and hematopathology.
• 00:42He has received many awards
• 00:43and recognitions, including being inducted
• 00:45into the American Society for
• 00:47Clinical Investigation.
• 00:49He also received the Pamela
• 00:51Catton Memorial Leukemia Research Award
• 00:53in twenty nineteen and the
• 00:55Ramsey Cotran Award from USCAP
• 00:57in twenty twenty two. He
• 00:58is currently an associate editor,
• 01:01of JCI,
• 01:02and he was recently appointed
• 01:04to the NIDDK
• 01:05Advisory Council.
• 01:07Since starting his independent lab
• 01:08in twenty eleven, doctor Gee
• 01:10has maintained continuous extramural support,
• 01:12securing twelve NIH grants. He
• 01:14is currently PI on three
• 01:15r o ones, an NSF
• 01:17award, two DOD grants, and
• 01:19fourteen foundation awards.
• 01:20A committed mentor,
• 01:22he directs Northwestern's
• 01:23pathology physician scientist training program
• 01:26and cofounded the Starzl Academy,
• 01:28guiding the next generation of
• 01:29clinician scientists.
• 01:31Today, we are honored to
• 01:32have him speak to us
• 01:32directly on this exciting story
• 01:34about the role of inflammation
• 01:36in the evolution from clonal
• 01:37hematopoiesis.
• 01:38Thank you.
• 01:45Thank you, Amina and Huapin
• 01:47and whole department,
• 01:49year pathology,
• 01:50for inviting me. It's really
• 01:52great great honor to be
• 01:53here.
• 01:54So,
• 01:55so today I'm just going
• 01:56to talk to you about
• 01:57the role of information in
• 01:59evolution from
• 02:00clonal hematopoiesis to MDS and
• 02:02AML.
• 02:04Before I start, I just
• 02:05have a quick disclosure. I'm
• 02:07a founder and co board
• 02:08members of two companies that
• 02:09I, co established.
• 02:12But today today's talk has
• 02:13nothing to do with that.
• 02:16Before I talk, really go
• 02:18into details about inflammation
• 02:19and myeloid disease,
• 02:21I just want to cover
• 02:23a little bit about what
• 02:24the major research is ongoing
• 02:26in my lab. So,
• 02:29ever since I started my
• 02:30own lab, back in two
• 02:31thousand twelve,
• 02:33I continue my postdoctoral work
• 02:35on universal poiseis. I was
• 02:37with Harvey Lodish,
• 02:39working on chromatin condensation
• 02:41and enucleation
• 02:42in rest cell development.
• 02:44So we have still our
• 02:45effort on doing in doing
• 02:47mechanistic studies in metapoisis
• 02:49as well as terminal erosive
• 02:50poiesis.
• 02:51Much of the effort on
• 02:52that part is, working on
• 02:54how
• 02:55using novel technologies including spatial
• 02:58omics studies to see
• 03:00to to tell the differences
• 03:01between mouse and human
• 03:03in hematopoiesis
• 03:04as well as urosopoiesis.
• 03:06The second major part is
• 03:08on myeloprolifera
• 03:09neoplasm. We focus on this
• 03:11protein called PLAGSTRING two,
• 03:13which is really a downstream
• 03:14target of JAK STAT5.
• 03:16And this protein is highly
• 03:18upregulated in patient with myeloproliferative
• 03:20neoplasm as well as in
• 03:22most models,
• 03:23with JAK2 mutation.
• 03:25So we are,
• 03:27we figure out how it
• 03:28works as a com o
• 03:30con, sacralin complex,
• 03:32activate AKT pathway, and we
• 03:35develop a small molecule,
• 03:36inhibitor of plaque two,
• 03:38to treat, hopefully to treat
• 03:40myelodysplast
• 03:40syndromes,
• 03:41myeloproliferative
• 03:42neoplasms.
• 03:44The third major part of
• 03:45my research lab, which is
• 03:47I'm going to talk about
• 03:48in detail today, is inflammation
• 03:50in innate immune signaling in
• 03:51myeloneoplasms.
• 03:54Major model, we use,
• 03:56mouse models,
• 03:57to mimic MDS as well
• 03:59as,
• 04:01bone marrow organoid models,
• 04:03and also human human's data,
• 04:06as well.
• 04:08So,
• 04:10so talking about myeloid diseases
• 04:12as well as inflammation, so
• 04:14we're really talking about,
• 04:16some of the disease or
• 04:17entities,
• 04:19that, listed here. So we're,
• 04:21so starting from non clonal
• 04:23idiopathic
• 04:24cytopenia of unknown significance,
• 04:27these kind of,
• 04:29we we don't call it
• 04:30disease, but this kind of,
• 04:32situation,
• 04:34where we cannot call them
• 04:35patients, but individuals will have
• 04:38unknown,
• 04:39cytopenias,
• 04:40and they don't have any
• 04:41detectable mutations.
• 04:44And these patients oftentimes, they
• 04:45can,
• 04:47they they can also have
• 04:48can further develop into
• 04:52chronic,
• 04:52so called clonal hematopoiesis of
• 04:54different inter determined potential.
• 04:57And in this patient, you
• 04:58can also you can you
• 04:59can also have,
• 05:01with or without cytopenia,
• 05:03but,
• 05:03oftentimes these patient, these individuals
• 05:06have,
• 05:07mutations detectable
• 05:08that I'm gonna talk about
• 05:10in detail later on.
• 05:11And then we have, entities
• 05:13called the clonal cytopenia on
• 05:15determined potential significance.
• 05:17And in these individuals,
• 05:19we not only have set
• 05:20opinions as well as have,
• 05:23mutations
• 05:24in many genes that I'm
• 05:25gonna talk about later on.
• 05:27So these patients all can,
• 05:30many of them can further
• 05:32progress to MDS,
• 05:34that's myelodysplastic syndrome.
• 05:36And we have low blast
• 05:38MDS and high blast MDS,
• 05:40and that's still based on
• 05:41previous WSL,
• 05:43classifications.
• 05:45And in these patients,
• 05:47most of them will have
• 05:48mutations or chromosome abnormalities.
• 05:51And, of course, they have
• 05:52bone marrow abnormalities with dysplasia
• 05:54in different kinds of lineages,
• 05:57as well as different percentage
• 05:59of, blasts
• 06:01that gives them different entities.
• 06:04And then, eventually,
• 06:06many of those patients can
• 06:08progress to acute myeloid leukemia
• 06:10where they have more than
• 06:11twenty percent of blasts either
• 06:13in the profile blood or
• 06:14in bone marrow.
• 06:15And they oftentimes,
• 06:17share similar mutations or chromosome
• 06:19abnormalities,
• 06:20as those patient with MDS.
• 06:24So,
• 06:25across all these, different entities,
• 06:27inflammation really play a big
• 06:29role, and there are a
• 06:30lot of studies,
• 06:33has been reported,
• 06:34in different aspects of inflammation,
• 06:37that seem, you know, that
• 06:38plays a role in different
• 06:40entities.
• 06:41But,
• 06:42as I mentioned,
• 06:44these entities have a lot
• 06:46of
• 06:47commonly shared in between these,
• 06:49entities as well as acute
• 06:51myeloid leukemia.
• 06:52The most common ones are,
• 06:54these are all called clonal
• 06:56hematopoietic
• 06:57clonal hematopoiesis mutations.
• 06:59The most common ones are
• 07:00DMT three a and tattoo
• 07:02mutations,
• 07:03as you can see from
• 07:04this,
• 07:06review paper we recently published.
• 07:10And,
• 07:11these
• 07:12common mutations,
• 07:13for example, DMT three a
• 07:15and tattoo,
• 07:16are commonly related to epigenetic
• 07:18modification, but they are also
• 07:20closely related to inflammation and
• 07:22inflammatory changes in the bone
• 07:23marrow overall environment.
• 07:26So,
• 07:29and and,
• 07:30and we are talking also
• 07:31about MDS when these,
• 07:34clonal hematopoietic mutation, patient with
• 07:36these mutations for the develop.
• 07:39And this MDS is really
• 07:41a a clonal,
• 07:43disorder that arrived from the
• 07:45bone marrow.
• 07:46And you have,
• 07:48a group of symptoms including
• 07:51cytopenia, dysplasia, and major and
• 07:54these can all cause to
• 07:55infective hematopoiesis
• 07:58and increase development to, acute
• 08:00myeloid leukemia. So mainly we
• 08:02have three lineages as you
• 08:03may you you know that
• 08:04in in
• 08:05the bone marrow environment.
• 08:07We have a red cell
• 08:08lineage. If you have dysplasia
• 08:10in the red cell lineage,
• 08:11you will have anemia.
• 08:14And we also have,
• 08:16myelolinage,
• 08:17that give rise to white
• 08:18blood cells. So dysplasia in
• 08:20white, myelolinage
• 08:21will cause,
• 08:23reduction in the red in
• 08:24the white blood cell count,
• 08:25and then it can lead
• 08:26to frequent infection,
• 08:28as well as weaken the
• 08:29immunity.
• 08:30And reduction in the megakaryocytic
• 08:32lineage that can cause,
• 08:35the reduction in in in
• 08:37platelets count, and that can
• 08:38lead to easy bleeding and,
• 08:40bruising.
• 08:41So these are all important
• 08:42symptoms that we, should keep
• 08:44in mind,
• 08:45and when we try to
• 08:46model these diseases in using
• 08:48animal
• 08:49models or other,
• 08:51nonhuman models.
• 08:54So we got interested in
• 08:55this field,
• 08:58especially,
• 09:00this specific entity called a
• 09:02delta five q m d
• 09:03s where
• 09:04the long,
• 09:06long arm of chromosome five
• 09:07is commonly deleted.
• 09:09And there are a lot
• 09:10of
• 09:11genes
• 09:12on chromosome five, as you
• 09:14can imagine.
• 09:15And one of the gene,
• 09:17is called, so one of
• 09:19the gene is, microRNA that
• 09:21has been shown before to
• 09:22be involved
• 09:24in thrombocytosis
• 09:25and,
• 09:26hypo,
• 09:27lobulated omega kerasocytes in in
• 09:29MDS.
• 09:31We are also interest interested
• 09:33in a gene called d
• 09:35I p h one, which
• 09:36encode protein called m dye
• 09:37one. So the the reason
• 09:39I'm
• 09:39getting to the MDS field
• 09:41is because,
• 09:43EIPH one, which encode m
• 09:45dye one, is actually involving
• 09:47acting cytoskeleton,
• 09:49modification. So
• 09:51we have discovered previously in
• 09:53our rest of your field
• 09:54that m dye one is
• 09:55important for rest of rest
• 09:57cell nucleation
• 09:58as well as,
• 10:00later stage of terminal universal
• 10:02poiseis.
• 10:03But it turns out that
• 10:04this gene is located on
• 10:06chromosome five q. So I
• 10:07am I've become very interested
• 10:09in chromosome five q division
• 10:11in MDS.
• 10:12So what we actually find
• 10:14out, that was back in
• 10:16more than ten years ago,
• 10:18using MDI one or dmp
• 10:20d I d I a
• 10:21ph one knockout mouse model,
• 10:23we found that these mice
• 10:25with when they are eight
• 10:27when they are about one
• 10:28year for
• 10:29old, they develop MDS like
• 10:31symptoms,
• 10:33where we found that actually
• 10:35CD14
• 10:35is overexpressed
• 10:37in apparently overexpressed on granular
• 10:40sites.
• 10:40Usually, a CD fourteen is
• 10:42over, overexpressed in monocytes.
• 10:45But in this, in this
• 10:46MOS model, CD fourteen is
• 10:48upregulated in granulocytes,
• 10:50and CD fourteen is a
• 10:51coreceptor with Tollac receptor four.
• 10:54And as
• 10:55TR four is commonly involved
• 10:57in the immune,
• 10:58abnormally in the immune signaling
• 11:00pathway,
• 11:01this activation
• 11:02or overexpression of CD fourteen
• 11:04leads to,
• 11:05inflammatory changes in this mouse
• 11:08model in their bone marrow,
• 11:09and that can lead to,
• 11:12granulocytopenia
• 11:13as well as dysplasia
• 11:15in in a variety of
• 11:16different lineages. And this, study
• 11:18was, led by, Ganesha Keith
• 11:21Forsen. He's currently has his
• 11:23own actually, he's in the
• 11:24business field, and actually he's
• 11:26running his own company.
• 11:30So with that,
• 11:31Yang Mei took over the
• 11:33project,
• 11:33after Ganesha left the lab.
• 11:36What Yang did is that,
• 11:38as I mentioned to you,
• 11:41m MDI1 is involved in
• 11:43the upper regulation of CD14,
• 11:45that corecept
• 11:46coreceptor
• 11:47with, colac receptor four pathway.
• 11:50And it has been shown
• 11:51that miR-one forty six,
• 11:53forty six a that I
• 11:55showed you before in the
• 11:56cartoon,
• 11:57is also involved in the
• 11:59upregulation
• 12:00or overactivation of Tollac receptor
• 12:02four pathway.
• 12:04So,
• 12:06Yang actually decided to cross
• 12:08these two knockout mice and
• 12:10make double knockout mice
• 12:12to more closely mimic patients
• 12:14with MDS, five weeks five
• 12:16q deletion.
• 12:17So this is,
• 12:19the
• 12:20ResCell count hemoglobin
• 12:22and metacritic
• 12:23count,
• 12:26entities,
• 12:27in this individually
• 12:28knockout mice. As you can
• 12:30see, compared to the to
• 12:31the wild type mice,
• 12:33the neuromphoresisait
• 12:35knockout mice as well as
• 12:37m by one knockout mice,
• 12:38they they show some phenotype
• 12:40but not as severe,
• 12:41compared to the wild type
• 12:43mice.
• 12:45And when he,
• 12:46checked the double knockout mice,
• 12:49you you can see really
• 12:50severe, significant difference compared to
• 12:52the single knockout.
• 12:54These mice with aging develop
• 12:56severe,
• 12:57anemia
• 12:58with reduction in RBC count
• 12:59and hemoglobin count as well
• 13:01as HCT.
• 13:03And if you look at
• 13:04their survival, these mice really,
• 13:07really die of the disease,
• 13:11at around one one around
• 13:13one year old
• 13:14compared to the single knockout
• 13:16mice.
• 13:18And, he also checked
• 13:20the different cytokine expression,
• 13:23inflammation
• 13:23in the bone marrow, And
• 13:25he found out that r
• 13:26six specifically,
• 13:29r six as well as
• 13:30TFR four are specifically highly
• 13:33operated
• 13:34in patient in mice with
• 13:35double knockout versus two genes.
• 13:38And he also checked
• 13:40the r six level in,
• 13:42g g one g r
• 13:43one positive mac one positive
• 13:44granulocytes.
• 13:46He found that, in both
• 13:48the bone marrow and the
• 13:49spleen,
• 13:49the double knockout mice have
• 13:51finally expressed our six levels,
• 13:53compared to the single knockout.
• 13:56This is also true for
• 13:57TNFr file expression.
• 13:59So this is a really,
• 14:00you know, it makes sense
• 14:01that double knockout mice, because
• 14:04they cotarget
• 14:07the TNF- Tollak receptor four
• 14:08pathway, they have this high
• 14:10upregulation,
• 14:11highly inflammatory changes in their
• 14:13bone marrow that lead to
• 14:14the eventual
• 14:15MDS development.
• 14:19So,
• 14:19here's the model that we,
• 14:21this paper was published in
• 14:22twenty eighteen
• 14:24in leukemia.
• 14:25So with the, we propose
• 14:27that with aging,
• 14:29and aging will lead to
• 14:31more
• 14:33damaged social molecular patterns and
• 14:34passage of social molecular pattern
• 14:36accumulation
• 14:38in individuals or particularly
• 14:40in this mouse model. And
• 14:41if you have both of
• 14:43these genes knocked out, you
• 14:45will have,
• 14:47especially you have a,
• 14:49myeloid derived suppressor cells. They
• 14:51are very sensitive to this
• 14:52environment, to the loss of
• 14:54these two genes, and they
• 14:55will over secrete
• 14:57TFR file as well as
• 14:59as r six. And these
• 15:00two cytokines actually have different
• 15:02mechanisms
• 15:03in, in,
• 15:05blocking
• 15:06common or universal voices for
• 15:08so for r six, they
• 15:10will lead to upregulation
• 15:12of ROS,
• 15:13that lead to upregulation
• 15:14activation of caspase three and
• 15:16seven. So that leads to
• 15:17the, apoptosis
• 15:19cell test. But for tnFR
• 15:21alpha that,
• 15:22they target,
• 15:23the GATA one. And GATA
• 15:25one is, as you know,
• 15:27is critical
• 15:28transferring factor for erosapoisis,
• 15:30and that leads
• 15:31to ineffective erosapoisis,
• 15:34and that leads to defective,
• 15:36rest cell production.
• 15:37So overall, this, this,
• 15:40these two kind of cytokines
• 15:41lead to ineffective rosuquoiesis and
• 15:44lead to a lot of
• 15:44cell death, and that can
• 15:46refuse,
• 15:48damage to sort of molecular
• 15:49patterns. And this is kind
• 15:51of like a positive feedback
• 15:52loop that promote the progression
• 15:55of this disease.
• 16:00So,
• 16:01Yang,
• 16:02kind of continue with this
• 16:03project.
• 16:05What he actually,
• 16:07did that we actually missed,
• 16:10in the previous publication in
• 16:11leukemia,
• 16:13because these mice,
• 16:15when they are one year
• 16:16old, they kind
• 16:18of die very quickly of
• 16:19the disease.
• 16:21But, in the follow-up study,
• 16:23he, Young really carefully looked
• 16:25at those moribund mice. And
• 16:27he found that there's a
• 16:28lot of,
• 16:30fibrotic changes in these mice
• 16:32in the bone marrow, as
• 16:33well
• 16:34as, a really accumulation of
• 16:36the, blast looking,
• 16:38cells,
• 16:39in their bone marrow, environment.
• 16:41And these, blast looking cells
• 16:43can also be found in
• 16:45the peripheral blood.
• 16:48So, we just,
• 16:50wonder whether because IL six
• 16:52and TRF are highly upregulated
• 16:54in the in this patient
• 16:55in this,
• 16:56in this MOS model, we
• 16:58just wonder whether r six
• 17:00particularly
• 17:01is critical
• 17:02to mediate this process.
• 17:04So, again,
• 17:05this is,
• 17:07the,
• 17:09all type comparison to double
• 17:11knockout
• 17:12of MDI1
• 17:13and miR46a.
• 17:15Again you can see dramatic,
• 17:17reduction
• 17:18in RBC, hemoglobin,
• 17:20as well as significant,
• 17:22thrombocytopenia
• 17:23in these mice when they
• 17:24are moribund.
• 17:26And then
• 17:27what he did is,
• 17:29on top of that, he
• 17:31additionally knocked out the r
• 17:32six. So he made triple
• 17:34knockout mice.
• 17:36So we thought at originally
• 17:38when we designed this experiment,
• 17:40because our supposed r six
• 17:41and t r four are
• 17:42regulated,
• 17:44probably r six knockout by
• 17:46itself may not be able
• 17:47to completely rescue the disease
• 17:49phenotype. But to our big
• 17:50surprise, this really dramatically
• 17:53the knockout of, R6 really
• 17:55dramatically rescued the disease phenotype.
• 17:57As you can see that,
• 18:00RBC
• 18:01goes back to almost like
• 18:02normal.
• 18:04Hemoglobin
• 18:05is normalized.
• 18:06Platelets is still a little
• 18:07bit low, but, it's really
• 18:09dramatically improved.
• 18:11Monocyte, which is,
• 18:13also highly upregulated in these
• 18:15double knockout mice, are also,
• 18:18their level also dropped down
• 18:19with, knockout to r six.
• 18:23And looking at their spleen,
• 18:24you can see that there
• 18:25is a this is a
• 18:26huge spleen in the double
• 18:27knockout mice. This is probably
• 18:30the the the biggest spleen
• 18:31I have ever seen in
• 18:32my in my research career.
• 18:35But
• 18:36knock out of our six
• 18:37completely
• 18:38normalized the spleen size,
• 18:40and their survival is also
• 18:43dramatically,
• 18:44improved.
• 18:47So, you know, I'm a
• 18:48hematopathologist,
• 18:49so I like to look
• 18:50at the morphology.
• 18:51So again, when we look
• 18:53at the double knockout mice,
• 18:54you can see that,
• 18:56again, these are moribond mice
• 18:58about to die.
• 18:59They are bone marrow is
• 19:00completely
• 19:01if,
• 19:02kind of infiltrated with these
• 19:03blast looking cells, including, you
• 19:05know, apoptotic bodies as well
• 19:07as mitotic bodies.
• 19:09There's
• 19:10no basic,
• 19:11differentiated,
• 19:12normal differentiated,
• 19:14cell types. For example, what
• 19:16you can see in the
• 19:17bowel type mice as well
• 19:19as our six knockout mice.
• 19:20It's nice megakaryocytes
• 19:21as, as well as these,
• 19:23granulocytes
• 19:24and the universal cells. They
• 19:26are completely,
• 19:27occupied by blasts.
• 19:29And if you have r
• 19:30six knockout,
• 19:32they are
• 19:33the the this phenotype is
• 19:35completely reworked.
• 19:37You can you you have
• 19:38megakaryocyte
• 19:39back into the shape. You
• 19:40have all these, segmented neutrophils.
• 19:43Actually, in neutrophils in mice
• 19:44are different from human. They're
• 19:46kind of like donut shaped.
• 19:49So you have you have
• 19:50full differentiation of different lineages.
• 19:53And the my and the
• 19:54fibrosis is also, back to
• 19:56the,
• 19:58is also significantly reduced.
• 20:00Also, spleen size, as shown,
• 20:02they are significantly reduced, and
• 20:03their, normal morphology where you
• 20:05can see both the red
• 20:07pulp and white pulp
• 20:09are also normalized with the
• 20:11knockout of r six.
• 20:15So all these are all
• 20:16happen,
• 20:17in most models. So what
• 20:18about human?
• 20:20So we actually,
• 20:22check the,
• 20:23published I think this is
• 20:24a form of one of
• 20:25the published dataset where we
• 20:27look at r six
• 20:29expression
• 20:30in different subtypes of MDS,
• 20:32including,
• 20:34again again, these are still
• 20:35previous,
• 20:36entities that from the previous
• 20:38WHO and, classification.
• 20:41So when you look at,
• 20:43compared to the control,
• 20:44healthy individuals,
• 20:46MDS with brain cellophiles have,
• 20:48not much upregulation of IL
• 20:50six receptor.
• 20:52MDS with single lineage dysplasia,
• 20:54not much, but MDS with
• 20:56those high risk MDS.
• 20:57MDS with excess plus one
• 20:59as well as plus two,
• 21:01have dramatically statistically significant
• 21:03upregulation of r six receptor.
• 21:06And this is, this is
• 21:07the correlate with their worst,
• 21:09survival,
• 21:11with increased r six receptor
• 21:13expression.
• 21:14So we we can also
• 21:15do,
• 21:17immunohistochemical
• 21:18stain,
• 21:19in these patients.
• 21:20So,
• 21:21with r six receptor, you
• 21:23can see that, for in
• 21:25both
• 21:26MDS with
• 21:27multilinear dysplasia, those are also
• 21:30high risk MDS
• 21:31as well as, MDS with
• 21:33excess plus.
• 21:34There are six receptor level
• 21:36are highly upregulated
• 21:37compared to the control ones,
• 21:39and
• 21:40and and there's a mildly
• 21:41upregulated out,
• 21:43r six receptor in MDS
• 21:44with rinsuloplasts.
• 21:47There's also a particular,
• 21:49subtype of r six receptor
• 21:51that's soluble.
• 21:52And we wonder whether this
• 21:54soluble,
• 21:56isoform of IR six is
• 21:58particularly upregulated in those MDS
• 22:00patients.
• 22:01And that's indeed the case,
• 22:03but, the upregulation of IR
• 22:05six, soluble IR six receptor
• 22:07is not something that you
• 22:08can see in the peripheral
• 22:09blood. But only if you,
• 22:11harvest the bone marrow solution,
• 22:13you can see the significant
• 22:15upregulation
• 22:16R6 receptor. And this also
• 22:18closely correlate with their disease,
• 22:21stage.
• 22:22Again, high risk MDS have
• 22:24much higher,
• 22:25or statistically significant upper regulation,
• 22:28soluble R6 receptor.
• 22:32So with that, we wonder
• 22:33whether,
• 22:35you target R6 receptor or
• 22:37R6 pathway can really,
• 22:40provide some therapeutic,
• 22:42effect
• 22:43in patient with MDS.
• 22:45So we actually took the,
• 22:48four patient samples,
• 22:50and this is a colony
• 22:51assay. And we treat the,
• 22:54these,
• 22:55CD thirty four derived,
• 22:57bone marrow colonies,
• 22:59using
• 23:00tocilizumab,
• 23:02which is our six receptor
• 23:03antibody.
• 23:05I think our six antibody,
• 23:07and that that can significantly,
• 23:09lead to the reduction specifically
• 23:11in MDS patient samples, but
• 23:13not in the normal CD34
• 23:15positive
• 23:16colonies.
• 23:17So this again demonstrate that,
• 23:19this, is I think these
• 23:20are all from high risk
• 23:21MDS patients. So these high
• 23:23risk MDS
• 23:24samples are very sensitive to
• 23:26our six receptor
• 23:27pathways.
• 23:30So with that, I just
• 23:31want to,
• 23:34summarize this part of my
• 23:35talk. So I think I
• 23:37I showed you that, double
• 23:38knockout mDY one as well
• 23:40as miRIF forty six a,
• 23:42causes age related to anemia
• 23:44and ineffective rosuploidosis
• 23:46and, damage associated molecular patterns
• 23:49that are increased
• 23:50in overaging.
• 23:52That that actually happens to
• 23:53everybody, not just MDS patients.
• 23:56That can trigger,
• 23:57the overproduction of r six
• 23:59and t alpha
• 24:00in in in these MOS
• 24:01models.
• 24:03And we show that pathological
• 24:05levels of these, proinflammatory
• 24:07cytokines can induce ineffective erosuppoiesis.
• 24:10And r six play a
• 24:11pivotal role in MDS progression
• 24:13to acute myeloid leukemia.
• 24:15And I would think that
• 24:16targeting r six pathway could
• 24:18be a therapeutic
• 24:19effective,
• 24:20in treating high risk MDS
• 24:22when they progress to acute
• 24:24myeloid leukemia.
• 24:26So,
• 24:27so with that, we kind
• 24:29of more,
• 24:30looking at inflammation overall,
• 24:33in in myeloid diseases,
• 24:35And this is a really
• 24:37simplified cartoon showing the basic
• 24:39inflammatory
• 24:40pathways.
• 24:41So once you have the
• 24:42damage associated molecular pattern or
• 24:44passage associated molecular patterns, that
• 24:45can trigger the activation of
• 24:47the inflammasomes,
• 24:49both from an RRP3 or
• 24:51as well as ASC.
• 24:52And this, together with the
• 24:54procaspace one, this can lead
• 24:56to the activation of caspace
• 24:57one, procaspace one to caspace
• 24:59one. And the activated caspace
• 25:01one can cleave pro IL
• 25:03one beta
• 25:04as as well as a
• 25:05pro r eighteen
• 25:07and lead to the, the
• 25:08the mature r one beta
• 25:10and r eighteen. And these,
• 25:11proinflammatory
• 25:12cytokines
• 25:14can secret out of this,
• 25:15this inflammatory cell into the,
• 25:18into the environment through this,
• 25:22this gas seventy immediately pore
• 25:23formation. So this actually was
• 25:25a really exciting discovery,
• 25:28I think, around
• 25:30a decade or so ago,
• 25:32mainly driven by, Feng Shao's
• 25:34group in China.
• 25:36And, so what happens is
• 25:37that activated the caspase one
• 25:39can also cleave the full
• 25:40length gas from a d
• 25:41and that d should so
• 25:43once you have the n
• 25:44terminal
• 25:45cleaved gastrimin d by caspase
• 25:47one,
• 25:48they can form a really
• 25:50beautiful pore,
• 25:51on the,
• 25:53cytoplasm membrane, and that leads
• 25:55to the release of these,
• 25:57proinflammatory
• 25:57cytokines.
• 26:00And here is, another cartoon
• 26:01that really shows vividly
• 26:04how this pore was formed.
• 26:06And actually, the crystal structure
• 26:08of, this n terminal domain
• 26:10of,
• 26:11gastrinomy d has been resolved
• 26:14And they form once you
• 26:15have this, n terminal domain
• 26:16cleave, they kind of form,
• 26:20kind of line up closely,
• 26:22and form this,
• 26:24palm like structure. And they
• 26:25they insert the,
• 26:27plasma membrane,
• 26:29with all these fingers
• 26:31into the and definitely form
• 26:33a kind of a really
• 26:34a beautiful circle and a
• 26:36big hole on the plasma
• 26:37membrane so that all those
• 26:39cytokines can be released from
• 26:40the, cytoplasm.
• 26:44So, we are interested in
• 26:46whether gas SMG
• 26:48is involved in the whole
• 26:49MDS inflammation,
• 26:52situation.
• 26:53So what we actually this
• 26:54is
• 26:55a data we this has
• 26:57not been published, but, they
• 26:58said the data we accumulated,
• 27:01many years ago.
• 27:02And we,
• 27:03again, we look at the
• 27:05control patient as well as
• 27:06delta five q MDS,
• 27:08MDS with reinstereoblast,
• 27:10MDS with high risk, excess
• 27:12of last one. You can
• 27:13see that, gastrin d expression
• 27:16is also highly
• 27:18increased,
• 27:19compared to the control patient
• 27:21in different MDS patients.
• 27:23Seems like MDS, derrick five
• 27:25q, we also is a
• 27:26low risk MDS. It the
• 27:27level of, carcinoid are also
• 27:30high, compared to
• 27:32the patient with, MDS ring
• 27:34cellophilus.
• 27:36So here's a quantification.
• 27:38Again, these are from the
• 27:40one of the database
• 27:42shared by, Amit Verma from
• 27:44Albert Einstein College of Medicine,
• 27:47and we can see that,
• 27:48EV patient with EMDS EV
• 27:50one, EV two,
• 27:52as well as,
• 27:53not much on retinoblast,
• 27:54but so they have a
• 27:55really high expression of a
• 27:57CasaMed.
• 27:58And looking back to our,
• 28:00double knockout mouse model that
• 28:02I just described,
• 28:03their level of and they
• 28:05can you can see clearly
• 28:06cleaved and terminal Casa MD
• 28:09here,
• 28:09compared to the wild type
• 28:11mice. So that indicate that
• 28:13these really double knockout mice,
• 28:15and this is specifically from
• 28:17the disease model because it
• 28:19takes time for these mice
• 28:20to develop MDS.
• 28:22It takes about eight months.
• 28:23So these are really samples
• 28:25from eight months to one
• 28:26year old mice.
• 28:28They are they they are
• 28:29really inflammatory being indicated by
• 28:32this cleaved and terminal guesstimate
• 28:34expression.
• 28:37So, this project was taken
• 28:40by Doctor Kehaan Ren.
• 28:43He's actually,
• 28:45he is going to,
• 28:46establish his own research lab
• 28:48at SUNY Stony Brook early
• 28:50next year.
• 28:52What he is trying to
• 28:54figure out is how weather
• 28:55gas m d,
• 28:57is really involving
• 28:59MDS as I mentioned, but
• 29:01particularly using the MOS model
• 29:02I just described to you,
• 29:04the double knockout of MDI
• 29:05one and neuron forty six
• 29:07a.
• 29:09So,
• 29:10he started with a single
• 29:11cell or R and C,
• 29:13experiment. And you can see
• 29:15here that compared to the
• 29:16wild type,
• 29:18mice,
• 29:19bone marrow cells from the
• 29:20double knockout mice have really
• 29:22increased, expression,
• 29:24in this particular population, which
• 29:26is the,
• 29:28the green is, monocyte and
• 29:30macrophage population.
• 29:32Is
• 29:33is, first of all, the
• 29:35the expression level of SMD
• 29:36is increased. And second,
• 29:39the double knockout mice, they
• 29:40have expanded
• 29:42monocytic and megakarousid
• 29:44macrophage population.
• 29:46So that's very interesting because
• 29:48monocyte is, is commonly known
• 29:50as,
• 29:51as as a cell type
• 29:52that secrete a lot of
• 29:53inflammatory cytokines.
• 29:56So
• 29:57he took the same approach
• 29:59as we did before, with
• 30:01our six knockout.
• 30:02And what he did here
• 30:04is,
• 30:04again,
• 30:06crossing double knockout mouse mouse
• 30:08with,
• 30:09Gethsemane d knockout. So he
• 30:11made Gethsemane d
• 30:13and GSMD
• 30:14neuron forty six a and
• 30:16DHP one MDI one triple
• 30:18knockout mice.
• 30:20And he used a different
• 30:22approach,
• 30:23than before,
• 30:24our six model, where he
• 30:27got these mouse models and
• 30:28he transplant
• 30:30the either wild type of
• 30:31double knockout or triple knockout
• 30:32might,
• 30:33bone marrow cells into the
• 30:35lethally irradiated recipient mice.
• 30:38So you can have a
• 30:38lot of mice, at the
• 30:40same time. And he looked
• 30:41at the survival of these
• 30:42mice,
• 30:43and you can see that,
• 30:45with double double knockout really
• 30:47die quickly as we've shown
• 30:49before. And,
• 30:51sort of as we expected,
• 30:53guess SMAD knockout,
• 30:55just as our six knock
• 30:56out, really significantly rescued their
• 30:58lethality.
• 30:59And there is also significant
• 31:00rescue of red blood cell
• 31:01count.
• 31:03Platelet count not much, but
• 31:05also a significant rescue of
• 31:07the increased monocyte count.
• 31:11This this, study also,
• 31:14demonstrate that, this disease is
• 31:16a hematopoietic
• 31:16intrinsic because it's trans transferable
• 31:19to, you know, to the
• 31:20to different mouse.
• 31:25So we show that there's
• 31:26an expansion of this
• 31:28monocytic,
• 31:29microphage,
• 31:30population.
• 31:32So we look, more closely,
• 31:34using this flow cytometry assay.
• 31:37As you can see that
• 31:38in the double knockout mice
• 31:40in the bone marrow, you
• 31:41have, not only,
• 31:43a growth overall overall growth
• 31:45overall expansion of monocyte macrophage
• 31:47population.
• 31:48You can also see the
• 31:49accumulation of this immature
• 31:51where you have
• 31:54RY six g negative negativity
• 31:56and low expression of r
• 31:58RY six c, which is
• 32:00a monocyte marker. So this
• 32:01is a kind of immature
• 32:02monocyte population.
• 32:04It's particularly
• 32:05increased in in the in
• 32:06the double knockout mouse model.
• 32:08And this, phenotype is reverted
• 32:11with a knockout of gastronomy
• 32:13d.
• 32:14So,
• 32:14this is a quantified,
• 32:17of the flow flow data.
• 32:21So this is again the,
• 32:23single cell, RNA seq data
• 32:25so you can, you know,
• 32:27as you can as you
• 32:28can imagine, you can characterize
• 32:29a lot of different cell
• 32:30populations,
• 32:31not only hematopoietic,
• 32:33but also stromal cells as
• 32:35well as mesenchymal
• 32:38stem cell populations.
• 32:41So,
• 32:42so but interestingly,
• 32:44if you transplant this double
• 32:46knockout mouse model,
• 32:48bone marrow cells into the
• 32:49recipient mice,
• 32:51the changes are not just
• 32:53happening in the double knockout
• 32:55donor cells. These donor cells
• 32:57can also,
• 32:58remodel the recipients non hematopoietic
• 33:01bone marrow environment,
• 33:02where you can see that,
• 33:04as as expected, neutrophils from
• 33:07the donor double knockout mouse
• 33:09model, they have once you
• 33:10have knockout of SNMD, they
• 33:12have reduction of these inflammatory
• 33:14markers,
• 33:15for example, s s one
• 33:16hundred and eighty nine,
• 33:18as well as the urethral
• 33:19population.
• 33:20But the in the fibroblast
• 33:22population as well as the
• 33:23mesenchymal stromal cells where they
• 33:25do not have these kind
• 33:26of genetic
• 33:28manipulations,
• 33:29their levels are also
• 33:32their level s one hundred
• 33:33and eighty nine levels are
• 33:34also decreased.
• 33:35So that's that's an interesting,
• 33:37finding from this model.
• 33:42So,
• 33:43so that also leads us
• 33:44to wonder whether
• 33:46what if you knock out
• 33:47the guesstimate in the bone
• 33:49marrow microenvironment?
• 33:50Would that also can,
• 33:52you know, revert the disease
• 33:53phenotype?
• 33:54So that's what he did.
• 33:56So he, again, he took
• 33:57the wild type mice,
• 34:00wild type donor cells,
• 34:02this oh, sorry.
• 34:04He he he took the
• 34:05top knockout mice and transplant
• 34:07them into wild type, either
• 34:08wild type or guesstimating knockout
• 34:11recipient mice. So in this,
• 34:13in these mice,
• 34:14SMD are completely knocked out
• 34:16in in every tissue type
• 34:18type,
• 34:19in in this in this
• 34:20mice.
• 34:21And then, actually, he, kind
• 34:23of, disappointed,
• 34:24that he didn't find any
• 34:26rescue of the of,
• 34:28the phenotypes
• 34:29as what we have shown
• 34:31before,
• 34:32using whole body guesstimate did
• 34:34not count. So that indicate
• 34:36that,
• 34:37probably
• 34:37not caught of GastroMD in
• 34:39the bone marrow microenvironment
• 34:40only
• 34:41may not be sufficient to
• 34:43rescue
• 34:44this disease phenotype because we
• 34:45thought that
• 34:46this, disease is quite strong
• 34:48because they can change from
• 34:49MDS to acute myeloid leukemia.
• 34:52And the inflammation is pretty
• 34:53pretty strong, so knockout of,
• 34:56or
• 34:57reduction in the inflammation in
• 34:58the bone marrow microenvironment
• 34:59may not be sufficient
• 35:01to revert this disease phenotype.
• 35:04So, so we then turn
• 35:06to a more milder,
• 35:08model
• 35:13MOS model.
• 35:14And tattoo, I mentioned a
• 35:15little bit in the beginning,
• 35:16but we know that tattoo
• 35:18is, is evolving
• 35:20in in the
• 35:22demystylation
• 35:23of,
• 35:24of chromatin and where they
• 35:26change from,
• 35:27five mysocerosine
• 35:28to five hydroxyamysocerosine
• 35:30and that lead to eventual
• 35:32demystylation.
• 35:33And they are also involved,
• 35:35in showing multiple publications
• 35:38that can,
• 35:39you know, tattoo is can
• 35:40repress the secretion of different
• 35:42cytokines including r one beta
• 35:44and r six in different
• 35:46tissue types.
• 35:48So we just wonder
• 35:49if we do the same
• 35:51experiment
• 35:52and transplant tattoo knockout mice
• 35:54where people have shown well
• 35:56established that they develop,
• 35:59inflammatory,
• 36:01myeloid disorders
• 36:03in these mice.
• 36:04Whether we transplant these knock
• 36:06on,
• 36:07bone marrow cells into the
• 36:08water into water type or
• 36:10a gas and the knock
• 36:11on it to see if
• 36:12there's any differences.
• 36:14So that's what we did.
• 36:16We can see that you
• 36:17can see that, once you
• 36:19have tattoo transplanting to wild
• 36:21type, there's a
• 36:22as we expected,
• 36:23the spleen is huge because
• 36:25they have this inflammation,
• 36:28and the extra medullary erossoposis,
• 36:30that's how they lead that
• 36:31lead to the enlargement of
• 36:33spleen.
• 36:34So if you knock out
• 36:36guess if you transplant tattoo
• 36:37into Casem D, knock out
• 36:39bone marrow microenvironment,
• 36:41This isn't the the
• 36:43the splenomegaly
• 36:44is not there anymore.
• 36:46And looking at the bone
• 36:48marrow,
• 36:49you can see that if
• 36:50you transplant knockout tattoo knockout
• 36:52into the wild type bone
• 36:53marrow,
• 36:54wild type recipient,
• 36:56You can see this inflammatory
• 36:57changes as well as dysplasia
• 37:00kind of infiltration
• 37:01of, immature appearance cells. It's
• 37:03not the full blown,
• 37:05leukemia,
• 37:06but they have some dysplastic
• 37:08changes.
• 37:09You know, most
• 37:10morphology is a little bit
• 37:11difficult compared to a human
• 37:13morphology, but I can see
• 37:14all I can see is
• 37:15there's
• 37:16a there's a, ineffective
• 37:18hematopoiesis
• 37:18in this in this model.
• 37:20And you can see splenomegaly
• 37:22as well as disruption of
• 37:23normal
• 37:24spleen architecture
• 37:26when you have
• 37:27tattoo knockout transplant into wild
• 37:29type recipient.
• 37:31This is,
• 37:32I would say,
• 37:34predominantly
• 37:35reverted
• 37:35when the tattoo knockout mice,
• 37:37bone marrow cells are transplanted
• 37:39into gastronomy d knockout bone
• 37:41marrow recipient,
• 37:42recipient mice.
• 37:47So,
• 37:48and when we look at
• 37:49the flow cytometry,
• 37:51you can see, as we
• 37:52expected,
• 37:53our six g,
• 37:55negative and our c our
• 37:57y six c molecule
• 37:59monocytic population
• 38:00that are increased with tattoo
• 38:02transplantation to water recipient
• 38:04are reduced,
• 38:05when they transplant into guesstimating
• 38:07knockout mice,
• 38:09both in the bone marrow,
• 38:11in the peripheral blood as
• 38:12well as in the bone
• 38:13marrow.
• 38:13CD eleven b,
• 38:15granulocytic,
• 38:16cells,
• 38:17myeloid cells as well are
• 38:19also reduced,
• 38:20when you transplant into guesstimating
• 38:22knockout.
• 38:24B cells that are re
• 38:26repressed when tattoo are transplanted
• 38:28into WALTAP
• 38:29are, are recovered when they
• 38:31transplant into GSMD knockout.
• 38:33And the c especially, significantly,
• 38:35CD seventy one that represent
• 38:39extra medullary erossopoises,
• 38:41the expansion of erossoprogenerate
• 38:43populations
• 38:43in the spleen,
• 38:45remarkably
• 38:46reverted when they transplanted into
• 38:48Casimod knock out.
• 38:51So this this data demonstrate
• 38:53that SMD,
• 38:54knock out of SMD in
• 38:56the bone marrow microenvironment
• 38:59can significantly
• 39:00revert the inflammatory changes or
• 39:02tetra mediated myeloid diseases,
• 39:05when,
• 39:07in this particular mouse model.
• 39:09So we also see that,
• 39:11the different cytokines,
• 39:13different side,
• 39:14inflammatory pathways
• 39:15are also upregulated.
• 39:17One tattoo are transplanted into
• 39:19wild type group,
• 39:21but, I don't see have
• 39:22data here, but I these
• 39:23changes are kind of reverted
• 39:25when they are transplanted into
• 39:26calcium D knockout group.
• 39:31So we can also,
• 39:33really,
• 39:34look at this,
• 39:36through a microscope,
• 39:38by doing immunofluorescence.
• 39:40And this is a
• 39:44confocal immunofluorescence
• 39:45study,
• 39:47and where you can see
• 39:48what we we stained the
• 39:49gas MND in N terminal
• 39:51domain. This is a specific
• 39:52for N terminal domain that
• 39:53indicate the inflammation in the
• 39:55bone marrow. We also stained
• 39:57CD two seventy one's,
• 39:59mesenchymal stromal marker,
• 40:01as well as UEA one
• 40:02that's also a, I think
• 40:04the it's,
• 40:05also a stromal cell marker.
• 40:07You can see that,
• 40:10water to water type, there's
• 40:12not much upregulation of guesstimating
• 40:14n terminals.
• 40:15But tattooed to water type,
• 40:16you can see a whole
• 40:17bone marrow is inflamed. It's
• 40:19kind of everywhere is has
• 40:20this red guesstimating n terminal
• 40:23deletion,
• 40:24deleted mutant,
• 40:26not mutant,
• 40:27isoform expression.
• 40:29But if you transplant
• 40:30tattoo knockout to GSMD knockout,
• 40:33model, recipient model,
• 40:35this inflammation is significantly
• 40:37reduced.
• 40:39Specifically, you can see the
• 40:41co, localization of the stromal
• 40:43markers
• 40:44with GSM d. So that
• 40:46indicate that
• 40:47it's not just the donor
• 40:49cells that with TAT2 mutation
• 40:51that are inflammatory.
• 40:53The bone marrow whole bone
• 40:54marrow environment is inflammatory.
• 40:56But with GASMAD knockout,
• 40:59you don't you don't have
• 41:00of course, you don't have
• 41:01GASMAD in the bone marrow
• 41:02microenvironment, but the the the
• 41:04the donor hematopoietic cells, their
• 41:06inflammation is also
• 41:10reduced. So,
• 41:12back to the single cell
• 41:13experiment,
• 41:15when we took the,
• 41:17bone marrow cells from these
• 41:18mice, from these transplanted mice,
• 41:20you can see that, as
• 41:22as expected, there's a outbreak
• 41:24significant outbreak of this inflammatory
• 41:28macrophages or monocytes
• 41:30when you transplant tattoo knockout
• 41:32to wild type, mice.
• 41:34But you can also see
• 41:35a significant outbreak ratio of
• 41:37CD four poly T cells,
• 41:39which is a reverted
• 41:41when tattoo
• 41:42knock out or transplanted to
• 41:43get SMD knock out mice.
• 41:45So this is interesting because,
• 41:49so I can I can
• 41:50go back to this slide,
• 41:52because you can you can
• 41:53see that, if we once
• 41:55we,
• 41:57took once we took this
• 41:59CD four polypropagation
• 42:00and further,
• 42:02some you know, subdivide them
• 42:04into different groups of CD
• 42:06four poly cells? You can
• 42:07see that compared to the
• 42:08wild type as well as
• 42:09get tattooed to guesstimate knockout,
• 42:11this particular population
• 42:13is is strongly upregulated.
• 42:15And this population expressed, of
• 42:17course, CD four, but also
• 42:19CD forty ligand and c,
• 42:21IL twenty one. So to
• 42:23us, this population,
• 42:25this CD four T cell
• 42:26population,
• 42:27express,
• 42:28t follicular helper cell like
• 42:30phenotype.
• 42:31So we are really thinking
• 42:32that,
• 42:34this could play a role
• 42:36in in kind of, communication
• 42:38between the,
• 42:40the CD4 poly T cells
• 42:42as well as
• 42:43the inflammatory macrophage and monocytes
• 42:46to to promote this co
• 42:47inflammatory
• 42:49bone marrow microenvironment.
• 42:51Back to this, data. So
• 42:53we also did,
• 42:55spatial transportomic studies. Here we
• 42:58use the 10x Visium platform,
• 43:01and you can see that,
• 43:03here is, so we spoke
• 43:05specifically looking at macrophages as
• 43:07well as, stromal cells. You
• 43:09can see that when you
• 43:10transplant tattoo knockout into wild
• 43:12type mice,
• 43:14there's a closer,
• 43:15not only there's a upregulation
• 43:17of, both,
• 43:19macrophages,
• 43:20of these macrophages,
• 43:22but these macrophages tend to,
• 43:25spatially,
• 43:27get closer or there is
• 43:28a space there is a
• 43:29spatial proximity
• 43:31of this inflammatory macrophages
• 43:33with the stromal cells. But
• 43:35if you use tattoo knockout,
• 43:38transplant into Casamine knockout,
• 43:41Of course, the microphage is
• 43:43reduced,
• 43:44and there's this,
• 43:45spatial
• 43:46proximity is not there anymore.
• 43:50So overall,
• 43:51we think this,
• 43:53this model like this.
• 43:55So we have operators in
• 43:57this,
• 43:59tattoo transplantation
• 44:00to to the wall to
• 44:02the not wall type recipient.
• 44:03You have, upper regulation of
• 44:05this inflammatory microphages
• 44:07and upper region for CD
• 44:08four poly T cells, where
• 44:10these CD four poly T
• 44:11cells,
• 44:12have CD,
• 44:14have high expression of CD
• 44:15forty ligand signaling,
• 44:17IR twenty one, which has
• 44:19been shown before that can
• 44:20induce macrophage proliferation.
• 44:23And macrophage these inflammatory macrophages,
• 44:26we we also have data
• 44:27showing that they also secret
• 44:29a lot of r one
• 44:30r twelve
• 44:31that can further enhance the
• 44:32expression or activation of the
• 44:34CD four positive T cells.
• 44:35So this kind of positive
• 44:37feedback
• 44:38really promoted the,
• 44:40the inflammation and disease progression
• 44:42of this particular model.
• 44:45And and overall,
• 44:47this, this is our overall,
• 44:49hypothesis.
• 44:50This This has not been
• 44:51published. We're still working,
• 44:54on really fine tune the
• 44:55different CD four part,
• 44:57cell populations and really try
• 44:59to figure out whether this,
• 45:02t follicular like helper cell
• 45:03population
• 45:04is truly
• 45:05really promote the,
• 45:07expansion of these, macrophages.
• 45:10And we can use some,
• 45:11you know, antibodies
• 45:13targeting
• 45:14CD forty ligand or r
• 45:15twenty one to demonstrate this
• 45:18pathway. So that's something,
• 45:20work in progress.
• 45:21But overall,
• 45:23so you you have this
• 45:24with with a tattoo mutation,
• 45:26you have this clonal hematopoietic
• 45:28population,
• 45:29that can, really
• 45:32have closer,
• 45:34spatial proximity to the wild
• 45:35type stromal cells
• 45:37and, whole and this whole
• 45:39bone marrow inflammatory environment,
• 45:42with the type two mutated
• 45:43microphases can really provide kind
• 45:45of positive feedback loop to
• 45:47promote the disease. But if
• 45:49you knock out guesstimate d,
• 45:51in this background,
• 45:52you kind of,
• 45:54disrupt this,
• 45:55positive feedback loop, and that
• 45:57leads to the reduction in
• 45:59the inflammatory,
• 46:00monocyte or macrophage population, and
• 46:03that leads to the also
• 46:04reduction in the CD4 populate
• 46:06positive population.
• 46:09So,
• 46:10again,
• 46:11this is still a working,
• 46:13work in progress model.
• 46:15So hopefully, we can once
• 46:17we get this antibody treatment
• 46:19experiment, we will be able
• 46:20to submit this manuscript by
• 46:22the end of this or
• 46:23early next year.
• 46:25So with that, I would
• 46:26like to thank people in
• 46:27my lab specifically,
• 46:29and as I mentioned, Yang
• 46:30Mei and Ke Han Ren
• 46:32really
• 46:32is the driving force of
• 46:34these these projects.
• 46:36And, other members of my
• 46:37lab, many of them are
• 46:39involved in,
• 46:40different projects including MDS, MTN,
• 46:44and spatialomic studies.
• 46:46And we also have a
• 46:46lot of,
• 46:48great colleagues in Northwestern,
• 46:50many people working
• 46:53on, different aspects of,
• 46:55myeloid diseases including Lucy Gottlieb,
• 46:58a collaboration
• 46:59on the germline mutated,
• 47:01MDS,
• 47:02models,
• 47:02Ian Liu, Liz Eklund.
• 47:05And we also have a
• 47:06a fantastic immunology group so
• 47:08where I can learn immunology
• 47:10from and inflammation from them,
• 47:12including Roman Ronan, Sumajin, Douyifang.
• 47:15And we also have a
• 47:17beautiful
• 47:18hematology,
• 47:19hematoposology
• 47:20group. Yihua Cheng is our
• 47:24program director, and Madina Suguna
• 47:26Hona is also collaborating on
• 47:28many of the
• 47:30projects I mentioned.
• 47:32And this is,
• 47:35so we also collaborated with,
• 47:37the drug discovery group at
• 47:39Northwestern.
• 47:40Many of them are involved
• 47:41in, you know, drug testing,
• 47:43as I mentioned, our six
• 47:44inhibit
• 47:45antibody testing
• 47:47as well as small molecule
• 47:48inhibitor development for the lab
• 47:51two study in MPN,
• 47:52and these are my funding
• 47:53resources.
• 47:55So I'll be happy to
• 47:56take any question you may
• 47:57have.
• 48:24So you're asking about r
• 48:26one signaling in your house
• 48:26slot. Can you put your
• 48:26way off? It's at work
• 48:26stands and aim here on
• 48:26our wall and stuff like
• 48:26that. So you're asking about
• 48:26r one
• 48:27signaling in this mouse model.
• 48:31So we, this model actually
• 48:33so based on our cytokine
• 48:34array data, r one is
• 48:36not particularly
• 48:37upregulated compared to you know,
• 48:39it's upregulated. It's the pan
• 48:41inflammatory cytokines are all upregulated.
• 48:43But compared to r six
• 48:45and t r alpha,
• 48:46r one is not dramatically
• 48:48high. So that's why we
• 48:49focused on r six and
• 48:50t r alpha instead of,
• 48:52r one beta. But r
• 48:53one has been studied in
• 48:55many other models
• 48:56in MDS as well as
• 48:57acute myeloid leukemia.
• 48:59It definitely play a big
• 49:01role in in this, pathogenesis
• 49:03myeloid diseases.
• 49:13Sorry. Okay? Do you know
• 49:14for the adaptive immune
• 49:16system?
• 49:18So far, that's where we
• 49:20are looking at. So we
• 49:22haven't really touched the adaptive
• 49:24immune system yet,
• 49:26but that's a good question.
• 49:29Whether
• 49:30b cell any types of
• 49:31b cells are involved,
• 49:32I don't know.
• 49:34Probably,
• 49:35but yeah. But I'm very
• 49:37interested in looking at the
• 49:39spatial
• 49:40omics studies
• 49:42in in these mouse models
• 49:43as well as patients to
• 49:44see their
• 49:46different adaptive immune system. In
• 49:47the immune system,
• 49:50there are any changes in
• 49:51the myeloid diseases,
• 49:53but not particularly in this
• 49:54model yet. And and they
• 49:56have already come in. I
• 49:57apologize.
• 49:58Is there
• 49:59a role
• 50:01for,
• 50:02program
• 50:04so that we basically wanna
• 50:06because everything here was, like,
• 50:07this this this is their
• 50:09update from a membrane.
• 50:13They
• 50:13assisted learning?
• 50:15Yes. Again, we haven't touched
• 50:17that yet,
• 50:18but it's possible. I mean,
• 50:19type two is, you know,
• 50:20commonly involving epigenetic and modifications,
• 50:24But that's, I think that's
• 50:25well known. So we we
• 50:27really didn't really touch specifically
• 50:29on epigenetic changes in this
• 50:31in
• 50:32this particular
• 50:33model system.
• 50:35Yeah.
• 50:45You see the effect in
• 50:46the tattoo not passed
• 50:48by limiting gas turbine in
• 50:50the host. Yeah. Right?
• 50:54Is as you expand those
• 50:56cells you put in, they're
• 50:57bringing with the wild type
• 50:59gas turbine into that environment,
• 51:01the transplanted cells.
• 51:03Do that does that play
• 51:04a role or link disease
• 51:06that become
• 51:08But the transplanted factor in
• 51:10and of itself. But the
• 51:11the diversity, what happens if
• 51:12you cannot help that strand
• 51:14and walk with that two
• 51:15and put into a wild
• 51:17type of person. That's that's
• 51:18the big thing.
• 51:19So
• 51:20the the transplanted cells are
• 51:22all tied to knockout. Right?
• 51:24Right. But they're gas terminus
• 51:25in wild. Yeah. Of course,
• 51:26they mean wild type. So
• 51:27that we have the data
• 51:28in, you know, this immunohoressence.
• 51:30They,
• 51:32with the knockout of the
• 51:33gastrin d, they suppress the
• 51:35expression of n terminal gasramin
• 51:37d in the transplanted
• 51:39mice,
• 51:40donor cells.
• 51:42Yes. Right. Right. But
• 51:44is to turn the experiment
• 51:45around, not have not just
• 51:47that tube, but also the
• 51:48gastric d in the
• 51:50transplant cells Yeah. Does that
• 51:54they are better when put
• 51:56into a wild type mice.
• 51:57So that won't that won't
• 51:58have disease, I I would
• 52:00say, because we we already
• 52:01did that with whole bone
• 52:02whole whole knockout mice. Right?
• 52:05So we we so that's
• 52:07that's what we showed.
• 52:09Yeah. So that's what we
• 52:10showed here.
• 52:14Here.
• 52:14So we did a triple
• 52:16knockout.
• 52:18Oh, sorry. You're talking about
• 52:20tattoo. Yeah. I would say
• 52:22it's will be similar to
• 52:23this. Similar to that? Yeah.
• 52:25So that's so even knocking
• 52:27out gas theremin in the
• 52:28cells,
• 52:31you don't need gas thermometer
• 52:32back out in the post.
• 52:34Right.
• 52:35Because that suppressed the intrinsic
• 52:38inflammatory changes.
• 52:39So they wouldn't induce the
• 52:43that's what I that's side
• 52:44of the equation works. Yeah.
• 52:47I think so. I think
• 52:48so. Yeah. So it play
• 52:50I think SMD play both
• 52:51intrinsic and extrinsic way role.
• 52:56That's what I'm like, we
• 52:57haven't done that experiment, but
• 52:59I think that's
• 53:01with this with this model,
• 53:02I think that's because this
• 53:03is a more stronger model.
• 53:05So I would I would
• 53:06imagine that tattoo knockout would
• 53:08be the same.
• 53:12Okay.
• 53:29Knockout? Oh. In that triple
• 53:31knockout? That's a good question.
• 53:33We haven't looked at that
• 53:34yet,
• 53:36but I would imagine because
• 53:40r six should still be
• 53:41upregulated,
• 53:43because
• 53:44not called r six.
• 53:46I don't know. It's not
• 53:47What happens to get? Yeah.
• 53:48Sturm is still upregulated. I
• 53:50mean, Destiny is still upregulated.
• 53:52It's not called the six.
• 53:53It's upregulated in all the
• 53:55activity
• 53:56that Gascam is doing is
• 53:57through y l six. I
• 53:59would imagine, but I I
• 54:00don't know yet,
• 54:02because we don't have that
• 54:03model.
• 54:05But that's a good question.
• 54:06Yeah.
• 54:09Mitra?
• 54:13I
• 54:18would think so. I would
• 54:19think so. There are there
• 54:21so we we also I
• 54:22didn't have data because we're
• 54:23still working on that. We
• 54:24use SMD inhibitors. There are
• 54:26different kinds of SMD inhibitors
• 54:29that can that has shown
• 54:31some effect in our most
• 54:32model, but we still find
• 54:34true that the datasets.
• 54:37Yeah. It's really great stuff.
• 54:39Thank you. My
• 54:41I really like this observation
• 54:42that
• 54:43the t follicular helper cells,
• 54:46the CD four t follicular
• 54:47helper software upregulated.
• 54:50That's how it comes. Right?
• 54:52And so those are gonna
• 54:53be active,
• 54:54you know, managing precipitation
• 54:56in the really in the
• 54:57dendritic cells
• 54:59primarily.
• 55:00So
• 55:01then that gets into this
• 55:02question.
• 55:06Can you, let's say, knock
• 55:07out sting just in the
• 55:09dead zone and
• 55:11modulate
• 55:13the ten to knock out?
• 55:15That's that's my question. Yeah.
• 55:17And I know that if
• 55:18I have made that maybe
• 55:18you don't wanna No.
• 55:22No. We we haven't done
• 55:23that yet. I think that
• 55:24would be something to think
• 55:25about.
• 55:26And the other question I
• 55:27I have related to that
• 55:29is,
• 55:30the,
• 55:32the gas and the phenotype,
• 55:34you know,
• 55:35it forms a four, I
• 55:37guess, in the cells. Right?
• 55:38Mhmm. And is that need
• 55:39to have a product cell
• 55:41debris? Is it does it
• 55:42kill the cells?
• 55:43Not really. It doesn't really
• 55:45so it depends on how
• 55:47how strongly gas in the
• 55:48internal domain is expressed, is
• 55:51cleaved. Right? So if you
• 55:52don't have enough gas in
• 55:53the that has been shown.
• 55:54That's not my major focus,
• 55:56but that has been shown
• 55:57by many publications.
• 55:59You have don't have enough
• 56:00gas in the n terminal
• 56:01domain.
• 56:02That cell inflammatory cells is
• 56:04not is is dying, but
• 56:06not truly die dead yet.
• 56:08Kind of like a zombie
• 56:09kind of stage. Is it
• 56:11release chromatin and nucleic acid?
• 56:15Could be. There is there
• 56:16I mean, if it does,
• 56:17right, that would suggest
• 56:20that a might be activated
• 56:21with the dead brain cells.
• 56:23I'm just trying to establish
• 56:24why
• 56:25Right. People living in their
• 56:26apple cells are increased. Yeah.
• 56:29It's so bay so what
• 56:31we hypothesize
• 56:32is that, the inflammatory
• 56:34monocytosis,
• 56:35microphages,
• 56:39provide this,
• 56:41kind of
• 56:42this has the highest r
• 56:44o twelve secondretion that has
• 56:46been shown to promote this
• 56:47t follicular help cell expansion.
• 56:50So that's our hypothesis,
• 56:52but we need to really
• 56:53show that experimentally.
• 57:12That yeah. That that could,
• 57:13yeah, that could also be
• 57:15the case. Yeah.
• 57:18Yeah. But that move back
• 57:19to the.
• 57:20We That's that's
• 57:22an interesting question. So we
• 57:23probably can make some generative
• 57:25cell specific,
• 57:27knockout
• 57:28model. In in these mouse
• 57:30models, have have you challenged
• 57:32them with an inflammatory signals
• 57:34like LPS or sepsis, like,
• 57:35conditionally?
• 57:37Right.
• 57:37Good question. But you don't
• 57:39need to. Yeah. I don't
• 57:40need to. So that that
• 57:41double knockout mouse model is
• 57:43very severe. I noticed the
• 57:44time time scale was months.
• 57:45And Yeah. But if you
• 57:47if you stimulate the LPS,
• 57:48do they just go down
• 57:49the next We we act
• 57:50we actually challenged the m
• 57:52by one single knockout
• 57:54in our original publication
• 57:56use using our PS. That
• 57:58can promote their disease.
• 58:00Because they may go down
• 58:01very fast because they're all
• 58:03primed for either metosis
• 58:05or
• 58:06metosis,
• 58:07which is you're having a
• 58:09macrophage
• 58:10enhancement in your and those
• 58:11are all prime all these
• 58:13signals are all the same.
• 58:14The IL one b Yeah.
• 58:15Gas derm and d for,
• 58:17macrophage extracellular
• 58:19traps too. And those they
• 58:21don't really kill the mouse.
• 58:22What happens is there's an
• 58:24occlusion somewhere else in a
• 58:25vital organ that causes organ
• 58:27failure. Nice die. Right. I
• 58:28wonder if you've been looking
• 58:29both of those,
• 58:31nettosis and metosis,
• 58:33or by citrullinated,
• 58:35of the histone Yeah. And
• 58:36if you did any work
• 58:38with a citrullinated
• 58:39antibody
• 58:40in your analysis. Yeah. Yeah.
• 58:43Yeah. That's work to be
• 58:44done. We haven't done that,
• 58:45unfortunately.
• 58:47Yeah. But that's, yeah. You
• 58:48can definitely
• 58:49I I don't remember if
• 58:51people have challenged
• 58:52the tattoo knock on. Must
• 58:54be, but that's a widely
• 58:55available model.
• 58:57I bet everything is primed
• 58:59for
• 58:59a catastrophic
• 59:01and vascular occlusions Right. From
• 59:04these extracellular Yeah.
• 59:08Yeah.
• 59:09Thank you so much. Thank
• 59:10you.