Canine Cancer Therapy: From Puppies to Patients

Transcript

• 00:00Like a blind email. He
• 00:02didn't know who I was.
• 00:03And about an hour later,
• 00:04I got a response.
• 00:07I got a response,
• 00:09and,
• 00:09he said, sure. I'll help
• 00:10if I can. Let's see
• 00:11if Willa, the dog with
• 00:13Willa's eligible.
• 00:14And within two weeks,
• 00:16the vaccine was on its
• 00:17way to Cincinnati to treat
• 00:18this dog. Willa was on
• 00:20her way to Cincinnati,
• 00:21had a leg amputated,
• 00:23got chemotherapy, and then got
• 00:24this vaccine that Mark was
• 00:25developing
• 00:26to treat dog panthers and
• 00:28dogs.
• 00:29And the last update I
• 00:30got about a year later
• 00:31was Willa
• 00:32was charging across this field
• 00:34with the three brothers,
• 00:36and I can guarantee she
• 00:37on three legs, she could
• 00:38outrun anyone in this in
• 00:40this room.
• 00:41So it's really a fantastic
• 00:42story. And Mark, as you'll
• 00:44hear, is now trying to
• 00:45use the same approach
• 00:47to treat patients, human patients,
• 00:49with these vaccines.
• 00:51And, even if it doesn't
• 00:52work in humans,
• 00:53at least he knows he's
• 00:54helping man's best friend. So
• 00:56that's something. So, Mark, thank
• 00:58you for coming.
• 01:03Thank you. It's terrific to
• 01:04be here. Thank you for
• 01:05the kind introduction. We're actually
• 01:07not treating patients yet, but
• 01:08if if you want to,
• 01:10I'd be happy to collaborate
• 01:12with
• 01:12what you're about to hear
• 01:13today.
• 01:16We've all been to,
• 01:18dozens or more of bench
• 01:20to bedside
• 01:23lectures.
• 01:24I got a little crazy
• 01:25with, alliteration
• 01:27here, obviously.
• 01:29Are there any
• 01:31dog owners
• 01:32here? Dog owners?
• 01:34Okay.
• 01:35Multiple dog owners. They have
• 01:37more than one. Okay.
• 01:39You,
• 01:40there are ninety million
• 01:43dogs in this country. Sixty
• 01:45million families with a dog.
• 01:47I have two for some
• 01:49one of those families that
• 01:51account for more than one.
• 01:53There are
• 01:54more dogs than kids under
• 01:56the age of eighteen
• 01:58in this country.
• 02:01I had to give a
• 02:03talk at the USDA, which,
• 02:05excuse me,
• 02:07regulates most of the veterinary
• 02:09biologics
• 02:10in this country. I had
• 02:11to remind them that,
• 02:12there are also more dogs
• 02:14than hogs in this,
• 02:17in this country, as well
• 02:18as more dogs than cows.
• 02:20So, only to emphasize the
• 02:22point that there are a
• 02:24whole lot of potential patients
• 02:26with potential cancers that we'd
• 02:28like to develop better therapies
• 02:30for. About
• 02:32one in four dogs in
• 02:33their lifetime
• 02:34will get a cancer.
• 02:36If your dog is lucky
• 02:38enough to live to ten
• 02:40years old,
• 02:41the chances of that dog,
• 02:44acquiring cancer are then about
• 02:45one in two.
• 02:48So today I'm gonna,
• 02:50of course, discuss,
• 02:52our studies and companion animals,
• 02:55try and make an argument
• 02:57that,
• 02:58those models
• 03:00well, actually they're not models.
• 03:01They're not models for human
• 03:03cancer at all. It's a
• 03:04parallel disease.
• 03:06And that's one point I'll
• 03:07try and emphasize today is
• 03:08that
• 03:09virtually
• 03:11all of the factors, the
• 03:12growth factors, mutations,
• 03:15predilection to metastases,
• 03:18therapies or therapeutic strategies are
• 03:21virtually identical. And you can
• 03:22I think all of us
• 03:23will agree that,
• 03:25abyss between
• 03:28treating mouse cancers and human
• 03:30cancers is far wider,
• 03:32than try treating, spontaneous
• 03:35dog cancers, which is what
• 03:37we're doing?
• 03:39I do have a conflict
• 03:40of interest,
• 03:41co founded a company that
• 03:43hopefully will manage,
• 03:45canine cancer care in the
• 03:47future.
• 03:50So this is my own
• 03:52pup that passed away about
• 03:53fourteen years ago of, inoperable,
• 03:56cardiac hemangiosarcoma.
• 03:58So it's,
• 04:00I've been both on the
• 04:02patient side, patient owner
• 04:04slot side, as well as
• 04:05now the therapeutic
• 04:07side. Hope you don't mind
• 04:08seeing
• 04:09a whole bunch of dog
• 04:10puppy pictures today.
• 04:13So
• 04:15what we're doing is an
• 04:17immune system based therapy. Of
• 04:18course, I have to make
• 04:19immunologists
• 04:20out of all of you,
• 04:21and this will take about
• 04:23ninety seconds, really. So,
• 04:26this is
• 04:27a diagram of a typical
• 04:30immune response to any anything,
• 04:32any foreign pathogen,
• 04:34any self protein or antigen,
• 04:37or even to tumors.
• 04:39So
• 04:40see if I can get
• 04:41this this oh, it is
• 04:43working. Okay. So it starts
• 04:44with an antigen presenting cell.
• 04:47In this diagram, it's dendritic
• 04:49cell. There are all sorts
• 04:50of antigen presenting cells, macrophages.
• 04:52B lymphocytes can present antigens
• 04:55very effectively, and I'll talk
• 04:56about that only briefly in
• 04:58the context of epitope spreading
• 05:00and
• 05:00why we think that may
• 05:02be important.
• 05:03But an antigen presenting cell
• 05:05does exactly that, presents small
• 05:08peptides on the surface
• 05:10of that dendritic cell or
• 05:11other APC in the context
• 05:13of major histocompatibility
• 05:15complex proteins.
• 05:17They typically first bump into
• 05:19T lymphocytes,
• 05:21that signal along with the
• 05:22second signal
• 05:23induces them or encourages
• 05:26them to develop into
• 05:27several different pathways.
• 05:29This is a very simplified
• 05:31version. You can either get
• 05:32CD eight, of course, restricted
• 05:34killer t cells that can
• 05:36kill pathogens or kill tumors
• 05:38directly, of course, Helper t
• 05:40cells, which do that, provide
• 05:42help to b cells. The
• 05:43end product
• 05:45being antibodies that bind that
• 05:47pathogen, hopefully clear it,
• 05:50or bind the tumor,
• 05:51hopefully clear it.
• 05:54Immunology makes a lot of
• 05:56sense
• 05:57to me. I think that
• 05:58may be why I went
• 05:59into it.
• 06:01The immune system responds when
• 06:02it needs to, when it
• 06:03sees something foreign
• 06:05and it's told
• 06:06to be turned off when
• 06:08that pathogen
• 06:09or whatever is triggering that
• 06:11immune response is cleared and
• 06:13gone.
• 06:15Okay.
• 06:16So,
• 06:19overall, I'm gonna talk about
• 06:20cancer neoantigens.
• 06:22Neoantigen just simply means a
• 06:23new antigen that your immune
• 06:25system has not seen.
• 06:30I neglected to mention that
• 06:32this process is also important
• 06:35in, quote, tolerance to lots
• 06:37of self issue proteins.
• 06:39So in your central immune
• 06:41organs like the thymus and
• 06:43bone marrow,
• 06:45Cell cells of the immune
• 06:46system, t cells primarily, will
• 06:48run into self proteins and
• 06:50be told to die.
• 06:51And about ninety plus percent
• 06:54of the t cells that
• 06:56develop in the thymus actually
• 06:58are specific for self proteins.
• 06:59They're typically
• 07:01told to die. Out in
• 07:02the periphery, they'll also bump
• 07:04into the self proteins and
• 07:05be tolerized.
• 07:06Again, this is to protect
• 07:08you from attacking your own
• 07:09tissues. Of course, the system's
• 07:11not perfect because
• 07:13we do have autoimmune diseases,
• 07:14and that's really
• 07:16represents,
• 07:17a flaw in this system
• 07:18of tolerance.
• 07:20But there are ways that
• 07:21that happen as well.
• 07:22So new antigens that your
• 07:24immune system
• 07:26can see and respond to.
• 07:28Now the cancer biologists
• 07:29typically think of neoantigens
• 07:32as
• 07:33products of mutation.
• 07:34You get a mutated gene,
• 07:36creates a novel self protein,
• 07:39a novel tumor protein that
• 07:40potentially
• 07:41the immune system can respond
• 07:43to. But I'm
• 07:45here to tell you that
• 07:46that's not the only type
• 07:48of neoantigen.
• 07:49Neoantigens can be
• 07:51any cryptic, any peptides of
• 07:54a lot of self proteins
• 07:55that just your immune system
• 07:56has never seen before
• 07:59can also be
• 08:01post translationally
• 08:02modified proteins.
• 08:04Proteins that come out of
• 08:05the translation pathway
• 08:07and get things like, glycosylation
• 08:10or phosphorylation
• 08:11or other many other,
• 08:13protein modifications
• 08:15that change the look
• 08:17of self proteins to your
• 08:18immune system.
• 08:20And
• 08:21regarding things,
• 08:24syndromes like autoimmune diseases and
• 08:26cancer,
• 08:27inflammation greatly amplifies the emergence,
• 08:31or the
• 08:32frequency
• 08:33of post translational modifications
• 08:35in tissues. So this is
• 08:36one primary way that both
• 08:39tumors via the tumor microenvironment,
• 08:41as well as other sites
• 08:42of inflammation,
• 08:44the pancreas,
• 08:45for example, and type one
• 08:47diabetes.
• 08:47Inflammation will crop will cause
• 08:49a number of different neoantigens
• 08:51to arise.
• 08:53How do you find them?
• 08:54Well, various omics, proteomics,
• 08:56of course, genomics.
• 08:58And what do we do
• 08:59with them once we find
• 09:00them? Well, we can think
• 09:01about using them as neoantigen
• 09:03therapies,
• 09:05either triggering immune responses to
• 09:07them or tolerance mechanisms.
• 09:09For example,
• 09:10allergens,
• 09:12that may be neoantigens.
• 09:14We can try and tolerance.
• 09:15We can try and shut
• 09:16down the immune system to
• 09:18neoantigens
• 09:19like those. They can also
• 09:21be used in diagnostics. I'll
• 09:23talk about that briefly in
• 09:24a minute.
• 09:26They can monitor,
• 09:28pathology of disease, be a
• 09:30marker of severity of pathology.
• 09:32I'll give you a few
• 09:33examples of that. So this
• 09:35is work that has come
• 09:36out of our laboratory in
• 09:37the last few years, just
• 09:38to emphasize the fact that,
• 09:40neoantigens
• 09:41are important in lots of
• 09:42syndromes like auto immune diseases.
• 09:45Notably,
• 09:45we found a protein modification
• 09:48in a pancreatic beta cell
• 09:51glucokinase.
• 09:52It's citrullinated
• 09:54Antibodies
• 09:55arise to that modified
• 09:56neoantigen
• 09:57in type one diabetes. And
• 09:59in fact, it's one of
• 10:00the earliest
• 10:01immunologic
• 10:02markers of type one diabetes.
• 10:05And we find it in
• 10:07human patients long before the
• 10:08onset of things like anti
• 10:10insulin antibodies.
• 10:11And it's becoming a diagnostic
• 10:14marker for diseases like,
• 10:16type one diabetes. Notably, rheumatoid
• 10:19arthritis is another disease that
• 10:21targets modified cell proteins, citrulline
• 10:23modified proteins. This is a
• 10:25diagnostic marker in the clinical
• 10:27labs upstairs.
• 10:31We did publish
• 10:33now a few papers about
• 10:34what I'll talk about today
• 10:35with some detail that, I
• 10:37won't be able to get
• 10:38to today. So you're welcome
• 10:40to look at those.
• 10:41And then I'll tell you
• 10:43some of our unpublished stories,
• 10:45today as well. So how
• 10:47do we find them? Again,
• 10:48either proteomics or genomics. This
• 10:50is one paper that I'll
• 10:51cite. There are many examples
• 10:54like this.
• 10:55You take a tumor sample
• 10:56from a patient,
• 11:00bring,
• 11:02you cleave
• 11:03proteins that are off the
• 11:04MHC molecule on immune cells,
• 11:08put them through mass spectroscopy,
• 11:10and identify
• 11:11whatever is sticking to those
• 11:13MHC proteins.
• 11:16Genomics, of course, define mutations.
• 11:18Those also predict,
• 11:20certain modified,
• 11:22neoantigens as well.
• 11:24The interesting thing though is
• 11:26that, clinically,
• 11:28at least,
• 11:29neoantigen specific t cells are
• 11:31linked with clinical efficacy of
• 11:33lots of other
• 11:35adjuvant therapies like checkpoint inhibitors.
• 11:38I'll talk a little bit
• 11:39in a minute about, the
• 11:41prominence of adoptive t cell
• 11:43therapies.
• 11:45It's not a perfect system
• 11:47in the laboratory, at least
• 11:48there are algorithms to predict
• 11:51neoantigens
• 11:51and when they're gonna bind
• 11:53to a particular MHC, those
• 11:54are not perfect. And in
• 11:56fact, a lot of the
• 11:58predicted
• 11:58neoantigens don't end up being
• 12:01a neoantigen
• 12:02at all.
• 12:04Once you get them, of
• 12:05course, you can do a
• 12:06number of things. You can
• 12:07choose to vaccinate against them
• 12:09in any number of ways.
• 12:10You can make mRNAs of
• 12:11those neoantigens,
• 12:13peptide
• 12:15based neoantigen therapies, which I'll
• 12:17talk more about today as
• 12:18well.
• 12:20This is one study out
• 12:22of non small cell lung
• 12:24cancer patients
• 12:26that show t cell responses
• 12:28to various neoantigens
• 12:29of KRAS and HER2.
• 12:32I bring it up because
• 12:34for a couple
• 12:35of interesting points is that
• 12:37based on
• 12:38the individual HLA
• 12:40makeup of individual patients, of
• 12:43course, they will respond to
• 12:44different neoantigens
• 12:46that's illustrated here. There are
• 12:48five patients that you can
• 12:49see here.
• 12:51And all you have to
• 12:53realize is that different t
• 12:55cells respond to different
• 12:57neoantigens
• 12:58in individual patients.
• 13:01The also,
• 13:02another point worth bringing up
• 13:04is in the boxes here,
• 13:05t cells
• 13:06are sometimes promiscuous.
• 13:09They will bind and respond
• 13:11to the neoantigen
• 13:12as well as to the
• 13:13native peptide, to the native
• 13:15unmodified protein.
• 13:17That's not unusual.
• 13:19So you really just don't
• 13:21know until you,
• 13:23experimentally
• 13:24determine,
• 13:25these outcomes.
• 13:31So
• 13:32in part, the reason,
• 13:34this kind of study is
• 13:35important, it also emphasizes the
• 13:37importance of epitope spreading
• 13:39in
• 13:40efficacy of,
• 13:42not only
• 13:43clearing pathogens, but of clearing
• 13:45tumors. And this is one
• 13:46recent study,
• 13:48illustrating
• 13:49that neoantigen therapies lead to
• 13:52this epitope spreading
• 13:53where not only one site
• 13:55on the target protein, tumor
• 13:57protein is bound, but also
• 13:59other sites
• 14:00on the tumor protein as
• 14:01well.
• 14:02That happens by a mechanism
• 14:04that, we studied now a
• 14:06few decades ago.
• 14:09In one manner in which
• 14:10this happens is that a
• 14:11B lymphocyte can be a
• 14:13terrific antigen presenting cell.
• 14:16Once that B cell is
• 14:17triggered by a neoantigen,
• 14:20even a short peptide,
• 14:22that b cell receptor often
• 14:23binds the native intact protein.
• 14:26So for example, this could
• 14:28be a protein that's a
• 14:30tissue protein in
• 14:32diabetes
• 14:33or cancer.
• 14:34It takes it up into
• 14:35the b cell.
• 14:37It digests that protein and
• 14:38presents
• 14:39now a number of different
• 14:41peptides on the surface
• 14:42for priming a second tier
• 14:45of t lymphocytes
• 14:47that in turn provide help
• 14:48to a second or third
• 14:49tier of b lymphocytes.
• 14:52So what was originally a
• 14:53very restricted
• 14:55immune response to a tissue
• 14:58antigen can become very diverse
• 15:00amplified and,
• 15:02that's what epitope spreading is.
• 15:04We know that that's important
• 15:06in effective cancer therapies.
• 15:11So,
• 15:15the media has
• 15:17been far ahead of actually
• 15:18many scientists in appreciating what
• 15:21studying various diseases in dogs
• 15:23can contribute to our understanding
• 15:25of human,
• 15:28pathology.
• 15:31So
• 15:32as I mentioned earlier is
• 15:34that, the canine cancer share,
• 15:37with almost
• 15:38complete identity, a lot of
• 15:40the tumor markers
• 15:41and pathways that human cancers
• 15:43do.
• 15:45You can see that humans
• 15:47and dogs do get virtually
• 15:49all the same types of
• 15:50cancer. Gliomas,
• 15:52oral melanoma
• 15:54in dogs actually a lot
• 15:55more frequent than you might
• 15:57appreciate.
• 15:58Liquid tumors,
• 16:00lymphomas and leukemias,
• 16:03breast cancers, bladder cancer, which
• 16:05is part of our own
• 16:06studies,
• 16:07prostate cancer,
• 16:08osteosarcomas
• 16:10are typically very frequent in
• 16:12dogs
• 16:13less. So
• 16:14it's a rare cancer in
• 16:16humans about a hundred thousand
• 16:17patients a year.
• 16:20So
• 16:21while the pathways
• 16:23are very similar regarding the
• 16:26genetics, the genomics, mutations,
• 16:29the predilection for metastases, even
• 16:31the sites of metastases in
• 16:33a particular cancer are virtually
• 16:35identical in in certain cancers
• 16:37in dogs and humans.
• 16:39The frequency is very different
• 16:41between dogs. That's one difference,
• 16:43between dogs and humans.
• 16:45And that's illustrated in the
• 16:46lower right. Dogs have a
• 16:48high frequency of liquid tumors,
• 16:50lymphomas, leukemias,
• 16:53soft tissue sarcomas,
• 16:55osteosarcomas,
• 16:56again, as I mentioned.
• 16:59There are a number of
• 17:01therapies that,
• 17:02have been translated
• 17:04from dogs to humans,
• 17:06including early attempts at bone
• 17:08marrow,
• 17:09transplantation,
• 17:10both autologous and allogeneic,
• 17:13various types of radiation therapy,
• 17:16limb sparing surgery. I'll have
• 17:19a illustration of that.
• 17:22Dogs make great models for
• 17:24doing
• 17:25drug,
• 17:26metabolism studies, pharmacokinetics.
• 17:30Those studies are typically
• 17:32much more accurate and a
• 17:34hundred pound dog than a
• 17:35twenty gram mouse,
• 17:37relative to what human,
• 17:39pharmacokinetics
• 17:40may find.
• 17:42And then certain drug therapies
• 17:44as well. Oncolytic viruses, I've
• 17:46listed just a few here.
• 17:47There have been a number
• 17:48of therapeutic,
• 17:51pathways for developing
• 17:53oncolytic viruses in canine cancers.
• 17:59Again,
• 18:01Limbs bearing surgery pioneered in
• 18:03dog surgery for
• 18:05osteosarcoma.
• 18:06This is from the early
• 18:08mid eighties.
• 18:09On the left,
• 18:11by comparison, a human cancer,
• 18:13human osteosarcoma
• 18:15patient with limb sparing surgery
• 18:17on the right.
• 18:20Human to canine, of course,
• 18:23the veterinary
• 18:25oncology communities
• 18:27use lots of drugs that
• 18:28are used off label treating
• 18:30dog cancers.
• 18:32Radiation therapy has been pioneered
• 18:33in humans,
• 18:35now applied to dogs.
• 18:38Checkpoint immunotherapies
• 18:40are only beginning
• 18:41to find their way into
• 18:43treating dog cancers. There is
• 18:45a USDA approved
• 18:46anti PD,
• 18:48one therapy made by Merck.
• 18:50We are collaborating with Merck
• 18:52in some of our studies,
• 18:54and there are anti PD
• 18:55l one inhibitors in process.
• 18:57And, of course, these
• 18:59need to be canonized in
• 19:01order to be used in
• 19:02in dog patients. The human,
• 19:05reagents just don't work because
• 19:06they're rejected.
• 19:08Lots of compounds, chemotherapies,
• 19:11that are now used again
• 19:12off label in treating dog
• 19:14cancers.
• 19:15Those are listed here.
• 19:19Lopatinib, which many of you
• 19:20know about small molecule inhibitors
• 19:22of both EGFR
• 19:24and HER2,
• 19:25one of the new first
• 19:27line treatments for bladder cancer
• 19:29in dogs.
• 19:33There's an interesting and useful
• 19:35for any of you doing
• 19:36genomics and wondering if your
• 19:39human genomics study have a
• 19:40parallel universe in dog cancers.
• 19:44There is a databank that
• 19:46you can access
• 19:48free of charge, of course,
• 19:51that's controlled by the NIH,
• 19:53and it's updated
• 19:55monthly,
• 19:56actually,
• 19:57that has lots of genomic
• 19:59studies and various dog cancer
• 20:01models.
• 20:02And this provides a source
• 20:04for
• 20:05data mining, if you will.
• 20:07If you're wondering if a
• 20:08particular human cancer mutation
• 20:11may be modeled in canine
• 20:12cancers, this is the perfect
• 20:14site to do that.
• 20:16In fact, it's been very
• 20:17helpful in defining,
• 20:19the genetics, the genomics, and
• 20:21molecular profiling,
• 20:24of gliomas
• 20:25and comparisons between human and
• 20:27canine gliomas.
• 20:30There's a terrific investigator that
• 20:32runs the comparative oncology program
• 20:34at the NIH,
• 20:35Amy LeBlanc. She manages this
• 20:38genomics library.
• 20:40She's, out of her own
• 20:42lab, defined
• 20:43mutations in
• 20:45a number of,
• 20:47genes comparable to human gliomas,
• 20:50p I three kinase, AKT,
• 20:52EGFR,
• 20:53t p fifty three, notably.
• 20:56There are similar
• 20:58methylation patterns in both canine
• 21:01and human
• 21:02gliomas.
• 21:03Overexpression
• 21:05of, platelet,
• 21:06derived growth factor alpha,
• 21:10EGFR
• 21:11amplification
• 21:12and various other mutations.
• 21:15And, her studies indicated that
• 21:18at least from the canine,
• 21:20glioma side is that those
• 21:21mutations
• 21:22more closely resemble
• 21:24pediatric
• 21:25gliomas as opposed to adult,
• 21:27which have,
• 21:28some very well defined differences.
• 21:33There's a second,
• 21:35group of studies done in
• 21:37osteosarcoma
• 21:38comparing human and canine.
• 21:40They share,
• 21:42frequently somatic copy numbers and
• 21:44alterations and mutations
• 21:46in TP fifty three
• 21:48and the other genes that
• 21:49you've can see here.
• 21:51They also have,
• 21:53a shared phenotype
• 21:55of the tumor microenvironment,
• 21:57meaning the cells,
• 21:58that,
• 22:00inhabit
• 22:01the primary sites of osteosarcoma
• 22:04comparable in both humans and
• 22:06dogs. Again,
• 22:07emphasizing the fact that what
• 22:09we learn about these,
• 22:12issues like tumor and microenvironment
• 22:14in dogs can greatly facilitate
• 22:16what we know and learn
• 22:18about the microenvironment
• 22:19in humans.
• 22:22So we picked the ERBB
• 22:25family of proteins to target
• 22:27neoantigens for a couple of
• 22:28reasons. One,
• 22:30is that we knew there
• 22:31are antibody based therapies that
• 22:34are effective against
• 22:35this family of proteins,
• 22:37Herceptin and Erbitux notably, of
• 22:39course.
• 22:41And I'm gonna tell you
• 22:42a story about that. These
• 22:43are, as you probably well
• 22:45know, these are surface signaling
• 22:48proteins found on,
• 22:50to a various extent on
• 22:52different tumors and even even,
• 22:55expressed differently
• 22:57in the primary site versus
• 22:59metastatic
• 23:00sites.
• 23:01We know from studying both
• 23:02human and canine
• 23:04cancers, for example, osteosarcoma
• 23:06metastases in the lung,
• 23:08express a very different profile
• 23:10of her B family proteins
• 23:13than is found in the
• 23:14primary site.
• 23:15And again, that triggers
• 23:17thoughts about how to treat
• 23:18this disease,
• 23:20creating therapies that may be
• 23:21effective against the primary site
• 23:23versus
• 23:24metastases.
• 23:27Canine studies
• 23:28have also you've been used
• 23:30to examine,
• 23:32why and how
• 23:33humans are refractory to various
• 23:36eGFR HER2
• 23:37drug therapies.
• 23:42So I'll just go over
• 23:43this briefly. This is, of
• 23:44course, the Erb family of
• 23:46proteins
• 23:47consists of EGFR,
• 23:48HER2, HER3, HER4.
• 23:50These are proteins that are
• 23:52found in monomers on the
• 23:54cell surface. They dimerize to
• 23:55create a signaling complex
• 23:58that signals within the cell
• 23:59goes to the nucleus and
• 24:02amplifies
• 24:03tumor cell proliferation,
• 24:05tumor cell survival,
• 24:08increases invasion and metastatic
• 24:10processes.
• 24:13There are ligands for the
• 24:15various ERB family members.
• 24:17And, of course, therapeutic strategies
• 24:19are designed
• 24:20at inhibiting
• 24:22these proteins either at the
• 24:23surface or within
• 24:25cells, of course.
• 24:26And this illustrates
• 24:27the human,
• 24:30both extracellular,
• 24:32extracellular,
• 24:33therapeutic strategies as well as
• 24:35intracellular
• 24:36ones. Cetuximab,
• 24:38for example,
• 24:39trastuzumab,
• 24:40Herceptin,
• 24:43as well as small molecule
• 24:45inhibitors of these signaling pathways.
• 24:47And I'm already mentioned that
• 24:48Lipidomid
• 24:49is,
• 24:51one of the new,
• 24:52drugs being treated,
• 24:54using to treat various canine
• 24:56cancers.
• 25:00So what
• 25:01canine cancers
• 25:03express this family of proteins?
• 25:05We have clinical trials ongoing
• 25:07in osteosarcoma,
• 25:08hemangiosarcoma,
• 25:10comparable to human angiosarcoma,
• 25:13transitional cell carcinoma,
• 25:15bladder cancer, if you will,
• 25:16in humans.
• 25:17And of course, a number
• 25:19of other dog cancers express
• 25:21various family members to various
• 25:23degrees, either eGFR HER2 or
• 25:26HER3. I'm not gonna talk
• 25:27about HER3 today
• 25:28too much.
• 25:31Notably, the liquid cancers do
• 25:32not express eGFR, of course.
• 25:36And it's not clear either
• 25:37in human studies or in
• 25:39our dog cancer studies how
• 25:41the level or the quantity
• 25:43of eGFR
• 25:44expression
• 25:45or how it may dimerize
• 25:48or heterodimerize
• 25:50on individual tumor cell effects
• 25:52or correlate to outcomes of
• 25:54various
• 25:55immune based therapies or even
• 25:56small molecule inhibitors.
• 26:00We were aided
• 26:02by
• 26:03studies that
• 26:05defined the crystal structure of
• 26:07eGFR.
• 26:08This is the extracellular
• 26:09domain
• 26:10of eGFR.
• 26:11And in red, you can
• 26:12see where cetuximab binds.
• 26:15In defining a potential neoantigen,
• 26:18we wanted to get at
• 26:19an extracellular
• 26:20site
• 26:21that was in close proximity
• 26:23of where cetuximab binds with
• 26:26the theory that potentially an
• 26:28immune response and antibody response
• 26:30to that site would have
• 26:31the same biology
• 26:32of,
• 26:33cetuximab.
• 26:36We picked one. You can
• 26:37see it here on orange.
• 26:39I'm not showing you the
• 26:40failures. We picked about six
• 26:42or eight or ten other
• 26:43sites that turned out not
• 26:45to be neoantigens
• 26:46at all and gendered or
• 26:48amplified no immune responses.
• 26:51And we first studied these
• 26:52in mouse models of cancer.
• 26:56So this is a site
• 26:57we ended up with.
• 26:59It is a site of
• 27:01high,
• 27:02almost identical homology between human
• 27:06and mice and dogs,
• 27:08and that is illustrated here.
• 27:10So
• 27:11human and mouse share exact
• 27:14amino acid sequence homology at
• 27:16this site that we chose.
• 27:19Dog,
• 27:20the dog site
• 27:22differs by one amino acid.
• 27:26It is also highly homologous
• 27:28to the site, a similar
• 27:30site on HER three and
• 27:32HER two
• 27:33that's,
• 27:34emphasized here.
• 27:36So that
• 27:38canine eGFR, HER2 and three
• 27:40all have a shared amino
• 27:42acid sequence
• 27:44on the surface of these
• 27:45proteins.
• 27:46And it was also surface
• 27:47accessible
• 27:49on these proteins as well.
• 27:51So the rationale here, which
• 27:53actually turned out to be
• 27:54true, is if that we
• 27:55can trigger
• 27:57an immune response antibodies and
• 27:58t cells that bind one
• 28:00site on eGFR that it
• 28:02may bind a homologous
• 28:04site on HER2 and HER3.
• 28:07And why is that important?
• 28:09Well,
• 28:11some forms of,
• 28:13mechanisms
• 28:14that control,
• 28:16inability to respond to eGFR
• 28:18therapies
• 28:19make humans refractory to various
• 28:22therapies
• 28:23are defined by those heterodimers,
• 28:26which ones heterodimerize
• 28:28with each other.
• 28:29So the rationale again is
• 28:31if eGFR
• 28:33chooses to heterodimerize
• 28:34with HER2 or HER3,
• 28:37that EGFR
• 28:38specific therapies
• 28:40may no longer work. But
• 28:41if one therapy, one antibody
• 28:43that could bind all three
• 28:44ligands,
• 28:47does that
• 28:49supersede
• 28:50that ability to,
• 28:52for tumor cells to resist
• 28:54killing by EGFR mediated
• 28:57immune therapies?
• 29:01So,
• 29:02these were and continue to
• 29:04be our exploratory
• 29:05studies.
• 29:06We're studying again three cancers,
• 29:07osteosarcoma,
• 29:08hemangiosarcoma,
• 29:09and bladder cancer.
• 29:11We typically
• 29:14ask that the veterinarians,
• 29:16in these clinical trials,
• 29:19this is a difficult question
• 29:21to, of course, answer is
• 29:22whether a dog or a
• 29:23human is going to survive
• 29:25three months or greater or
• 29:26longer. And the reason we
• 29:28did that is this is
• 29:29an immune based therapy.
• 29:31It's two injections, three weeks
• 29:33apart. It takes about three
• 29:35or four weeks to generate
• 29:37an active antibody or t
• 29:39cell response to these neoantigens.
• 29:42So
• 29:43in order to define efficacy,
• 29:45we wanted to at least
• 29:47give
• 29:47the patients, the dogs,
• 29:49a chance to make a
• 29:50vigorous immune response, and then
• 29:52we could hopefully measure some
• 29:54outcomes that were meaningful.
• 29:57So it's this eGFR,
• 30:00peptide neoantigen.
• 30:02We mix toll like receptor
• 30:04agonist,
• 30:05can be CPG. We've used
• 30:06others as well.
• 30:08And an oil based adjuvant
• 30:10called montanide ISA fifty one.
• 30:12It's made by a company
• 30:14in France.
• 30:15This is a GMP
• 30:17quality
• 30:18oil adjuvant. It's already used
• 30:21in a number of human
• 30:22based
• 30:23vaccination studies.
• 30:25So they get two injections
• 30:27three weeks apart. We take
• 30:29blood samples throughout.
• 30:31We
• 30:32assay them by flow cytometry
• 30:34and then note and many
• 30:36several other laboratory,
• 30:39processes.
• 30:40We try and get tissues,
• 30:41tumor tissues from dogs throughout.
• 30:44If we can get them,
• 30:45not always easy to do.
• 30:47For this study, radiographs,
• 30:50are required every three months.
• 30:51So we can potentially
• 30:52follow metastases
• 30:54of these patients.
• 30:56And, everything's uploaded,
• 30:58into,
• 30:59the Yale REDCap
• 31:01system, much like
• 31:03all the other clinical trials
• 31:04are here. So we have
• 31:07very well articulated
• 31:09data from our patient subsets,
• 31:12date of birth, date of
• 31:13diagnosis,
• 31:15every clinical visit, every radiograph,
• 31:19other meds that the dogs
• 31:21are on,
• 31:22gender,
• 31:23breed,
• 31:25Trying to think what else?
• 31:26What other things do we
• 31:27collect? Esther Renell.
• 31:30Am I getting most of
• 31:31them? Getting most of the
• 31:32important ones at least.
• 31:34Okay. So here's the overall
• 31:36strategy.
• 31:37Dog comes into the,
• 31:38clinic,
• 31:40already has pathology
• 31:41diagnosed
• 31:43tumors of those three types.
• 31:46They will get standard of
• 31:47care.
• 31:48And depending on the cancer
• 31:50type, standard of care, for
• 31:51example, for osteosarcoma,
• 31:54at least appendicular
• 31:55osteosarcoma
• 31:56is amputation.
• 31:58Some get,
• 32:01limb sparing surgery, very few
• 32:03anymore though.
• 32:05For this amputation and carboplatin,
• 32:07four to six rounds with
• 32:08or without our neoantigen
• 32:10therapy.
• 32:12There are other standards of
• 32:13care for the other tumors,
• 32:14which I'll talk about in
• 32:15a second. So again, the
• 32:17strategy, inject the dog twice.
• 32:19We can measure antibodies that
• 32:20arise to the neoantigen.
• 32:23And the strategy is that
• 32:24we're blocking
• 32:25or directly killing eGFR
• 32:28bearing tumor targets.
• 32:30So these are the serologies
• 32:32of several cohorts of our
• 32:34dog patients
• 32:36just like how humans respond
• 32:38to a particular vaccination,
• 32:41such as the same with
• 32:42dogs. Humans respond differently because
• 32:45we all have different HLA,
• 32:48composition.
• 32:49So we all don't,
• 32:51respond the same to things
• 32:53like flu or COVID vaccination.
• 32:55We will have different titers,
• 32:57different levels of immune responses,
• 32:59such as the case for
• 33:00dogs to this neoantigen.
• 33:03We get anywhere from four
• 33:04to thirty fold
• 33:06increases in,
• 33:08antibody responses to this protein.
• 33:13Importantly,
• 33:14the immune responses the antibody
• 33:15responses do bind,
• 33:18cell based eGFR
• 33:20when it's presented on living
• 33:22cells.
• 33:22These are a four three
• 33:24one human tumor cell lines
• 33:26that express eGFR.
• 33:29They also the immune responses
• 33:31also bind
• 33:33a HER2 bearing
• 33:35human tumor cell,
• 33:36MDA MB four five three.
• 33:38These do not express eGFR.
• 33:41Again, supporting
• 33:42the outcome that
• 33:44this cross reactive peptide, the
• 33:46sequence that is shared between
• 33:48eGFR
• 33:49HER2 and HER3
• 33:50does in fact
• 33:52bind the native protein on
• 33:53cells in which it resides.
• 33:59Okay. So then we went
• 34:01to really the strategies that
• 34:02were used to screen
• 34:05Herceptin and Erbitux when they
• 34:06were being developed.
• 34:08How do they kill tumors?
• 34:09Do they block
• 34:10signaling
• 34:11through the cell surface proteins?
• 34:13Do they kill tumors directly?
• 34:16This is
• 34:17one example of
• 34:19how immune responses in our
• 34:21dog patients block signaling
• 34:23through the eGFR
• 34:24and HER2 pathways.
• 34:26So for example, in the
• 34:28top panel, this is a
• 34:29canine
• 34:30osteosarcoma
• 34:31cell line.
• 34:32We can measure phospho eGFR.
• 34:35No antibody gives this signal.
• 34:38Anti EGFR
• 34:39control antibody blocks that signaling.
• 34:42Pre immune, again, not blocked.
• 34:44Immune serum from one of
• 34:46our dogs blocks very nicely
• 34:47nicely.
• 34:49Graphical representation
• 34:50of that is here.
• 34:52Also
• 34:53inhibits,
• 34:54signaling through a human,
• 34:57eGFR bearing cell line a
• 34:59four three one that's illustrated
• 35:01here.
• 35:02So, again, has the biology
• 35:04that you want in an
• 35:05eGFR
• 35:06therapy blocked signaling? One of
• 35:08the things you want.
• 35:13So do the immune responses
• 35:14actually bind tumor tissue? And
• 35:16this is a number of,
• 35:20staining
• 35:20for how that happens.
• 35:22There are a number of
• 35:23dogs listed,
• 35:25on this slide.
• 35:27Controls on the right, normal
• 35:28dog serum, and eGFR
• 35:31antibody control.
• 35:33Excuse me.
• 35:35Various dog
• 35:36sera from our cohort, either
• 35:38preimmune or immune sera.
• 35:41These are osteosarcoma
• 35:42cells lighting up with post
• 35:44immune serum. Again, indicating that
• 35:47those osteosarcoma
• 35:48cells are expressing the targets
• 35:50that are,
• 35:52induced by our neoantigen
• 35:54immunization.
• 35:59So
• 36:00getting to some of the
• 36:01other nuances, osteosarcoma
• 36:03is a disease both in
• 36:04humans and in dogs in
• 36:06which
• 36:07metastases
• 36:08to the lung
• 36:09is the most important
• 36:11factor of morbidity and mortality.
• 36:14This is one patient that,
• 36:16in fact, had that. This
• 36:17is Cody.
• 36:19Dog HIPAA,
• 36:20rules are very different than
• 36:22human. Cody didn't mind. I
• 36:23got his consent, actually.
• 36:25So Cody had amputation of
• 36:27a primary tumor,
• 36:29osteosarcoma,
• 36:30of course, left front leg
• 36:31as you can see,
• 36:33started to fail
• 36:35conventional therapy, carboplatin.
• 36:38At a metastasis to the
• 36:39lung,
• 36:40we enrolled this dog. I
• 36:42wasn't actually expecting much to
• 36:44happen.
• 36:44Within about six or eight
• 36:46months, that, lung metastasis
• 36:49resolved.
• 36:50Cody lived,
• 36:51another three and a half
• 36:52years.
• 36:54Ended up actually getting a
• 36:55second unrelated tumor, hemangiosarcoma,
• 36:58from which, the dog did
• 36:59not survive. So
• 37:03overall, among a cohort of
• 37:04osteosarcoma
• 37:06patients, specifically,
• 37:08standard of care twelve months
• 37:10survival with standard of care,
• 37:11amputation,
• 37:12carboplatin,
• 37:14about thirty to thirty five
• 37:16or forty percent of dogs
• 37:17will survive
• 37:18one year, twelve months.
• 37:21Adding
• 37:22this neoantigen
• 37:23therapy to standard of care,
• 37:25increases survival to about sixty,
• 37:28sixty five percent. So quite
• 37:30a dramatic difference. And that's
• 37:31taking all,
• 37:33all commerce, all
• 37:35dogs within this group with
• 37:37or without metastases. And I'll
• 37:39tell you more about that
• 37:40in a minute.
• 37:41And the panels on the
• 37:42right, again, just to show
• 37:43that antibodies from Cody bind
• 37:46both dog and human cell
• 37:48lines that, express these proteins,
• 37:51the ERB family of proteins.
• 37:55Cody was not a one
• 37:56off
• 37:57for clearing a lung metastases.
• 38:00We have at least four
• 38:01other examples.
• 38:04Three are on the top
• 38:05panels here.
• 38:06We
• 38:07see resolution or at least
• 38:09stasis of many,
• 38:11lung METs, meaning they don't
• 38:13change in size over time.
• 38:15These are three that happened
• 38:16to resolve,
• 38:17happens anywhere between three and
• 38:19eleven months after
• 38:20initiation of therapy.
• 38:22The bottom panel is a
• 38:23primary site in the hip
• 38:25that couldn't be removed surgically.
• 38:27But over now,
• 38:29close to two years,
• 38:31which,
• 38:32is still gone, still resolved
• 38:34in this particular patient.
• 38:37So again, in humans and
• 38:39in dogs, this is the
• 38:41biggest problem
• 38:42in survival,
• 38:44in this patient subset in
• 38:46this type of cancer.
• 38:49There we have an ongoing
• 38:50study at Washington State University,
• 38:53vet school.
• 38:54Rancellon is running that. Rance
• 38:56has a program for treating
• 38:57canine osteosarcoma
• 38:59without amputation,
• 39:01without chemotherapy.
• 39:02It's irradiation therapy,
• 39:05eight gray over given over
• 39:07two days,
• 39:08with or without our neoantigen
• 39:11eGFR
• 39:12therapy. And this is very
• 39:13recent data from France at
• 39:15Washington State.
• 39:16So in his cohort, the
• 39:18number is not great. Eleven
• 39:19patients,
• 39:21survival again, this is just
• 39:22radiation in EGFR neoantigen therapy.
• 39:27A median survival of almost
• 39:28a year
• 39:30without neoantigen therapy from his
• 39:33cohort of of patients, again,
• 39:35done right in his clinics,
• 39:38excluding our eGFR therapy patients
• 39:41only survive a hundred and
• 39:42thirty six plus or minus
• 39:44days.
• 39:45So, again,
• 39:47survival
• 39:48benefit with this neoantigen
• 39:50therapy.
• 39:55Overall, this is from different
• 39:57clinics, not Washington State University.
• 40:00This is, again, amputation,
• 40:03chemotherapy, carboplatin
• 40:04plus or minus eGFR therapy.
• 40:07All of these dogs did
• 40:08not have lung mets on
• 40:11original diagnosis.
• 40:13Their median survival is about,
• 40:15three eighty eight days, two
• 40:17years survival is thirty one
• 40:19percent. Again, significantly better than
• 40:22standard of care.
• 40:25We subset of these patients.
• 40:27Now, in canine
• 40:29osteosarcoma,
• 40:30virtually all patients
• 40:32will get a lung MET
• 40:34within about one year.
• 40:38In our study, these were
• 40:39all dogs that did not
• 40:41have lung METs at the
• 40:42origin of the study, got
• 40:44our therapy with standard of
• 40:45care. Only about half of
• 40:47them ended up getting lung
• 40:48mets. And this is the
• 40:49survival curve for those that
• 40:51either did
• 40:52get lung mets or remained
• 40:54metastasis
• 40:55free. As you'd predict, the
• 40:56ones that didn't get mets
• 40:58live significantly longer. Median survival
• 41:01now is well over a
• 41:02year versus
• 41:03survival,
• 41:05in dogs that did get
• 41:07lung mets of less than
• 41:08a year, two twenty nine
• 41:09days.
• 41:11So we're just now beginning
• 41:13or trying to understand how
• 41:14things like the tumor microenvironment,
• 41:16expression of various levels of
• 41:18eGFR
• 41:19on individual
• 41:20osteosarcoma
• 41:21patients
• 41:22may reflect,
• 41:24clinical efficacy
• 41:26or perhaps
• 41:27the magnitude
• 41:28of the immune response to
• 41:29this eGFR neoantigen therapy.
• 41:33Again, comparing human studies. Localized
• 41:36disease survive five year survival
• 41:38is pretty good, Seventy six
• 41:40percent. If there's regional spread
• 41:42in osteo human osteosarcoma
• 41:44less,
• 41:45as I mentioned earlier, METS
• 41:47to the lung,
• 41:48not good in humans either.
• 41:53Gonna move to hemangiosarcoma.
• 41:56This is arises in the
• 41:58spleen of dogs.
• 42:00Standard of care is splenectomy
• 42:02and doxorubicin
• 42:04with or without our eGFR
• 42:07therapy.
• 42:08So without are the yellow
• 42:09lines in the top panel.
• 42:12Stage one is localized disease
• 42:13in the spleen.
• 42:15Stage two has
• 42:16some
• 42:17infiltration of subcutaneous
• 42:19tissues in the spleen. And
• 42:21stage three has distant METs,
• 42:24liver or elsewhere.
• 42:26So
• 42:27significant improvement
• 42:28of,
• 42:30survival
• 42:31in dogs getting splenectomy,
• 42:33doxorubicin,
• 42:34and eGFR therapy.
• 42:37Two hundred and let's see.
• 42:39Two hundred and thirty six
• 42:40days. Hemangiosarcoma
• 42:42is a very aggressive dog
• 42:44cancer.
• 42:45Most dogs do not survive
• 42:47more than
• 42:48sixty days, ninety days at
• 42:50best.
• 42:51And that's even with surgery.
• 42:55Dog untreated dogs will rarely
• 42:57live longer than thirty days.
• 43:00Again, a statistically
• 43:02very significant
• 43:03approval of survival and stage
• 43:04two disease. Again, more infiltrative
• 43:07disease,
• 43:08compared to,
• 43:10dogs that do not get
• 43:12eGFR therapy.
• 43:13Stage three with METS
• 43:16couldn't do any good.
• 43:17Just extensive disease. We don't,
• 43:20we don't change the curve
• 43:22compared to standard of care.
• 43:25We were
• 43:26lucky enough to get a
• 43:27dog's spleen,
• 43:30that had already had our
• 43:32therapy. We simply
• 43:33use the tissue and asked,
• 43:35are there antibodies that are
• 43:36infiltrating
• 43:37that tumor tissue? The answer
• 43:39was yes. That's illustrated here
• 43:41on the right.
• 43:44C d eight t cells
• 43:45infiltrate those tissues as well.
• 43:47This is,
• 43:48a hemangiosarcoma
• 43:49tissue stained for c d
• 43:51a t cells.
• 43:52Again, after e g f
• 43:53r therapy.
• 43:55This is a normal Hemangiosarcoma
• 43:57tissue stained with pre immune
• 43:58and immune dog serum.
• 44:01Again, illustrating the brightness on
• 44:03the right of eGFR HER2,
• 44:06staining patterns.
• 44:09And then finally, bladder cancer.
• 44:11Again, very similar to human
• 44:13disease.
• 44:14Dogs don't do quite as
• 44:16well with it.
• 44:17We get a mixed,
• 44:20outcome of survival.
• 44:22Bladder cancer really does not,
• 44:24at least in dogs, does
• 44:25not have a perfect standard
• 44:27of care. Sometimes it can
• 44:28be surgically treated if it's
• 44:30appropriate, if it's not,
• 44:33infiltrated into tissue.
• 44:36Different types of bladder cancer,
• 44:38urethral bladder or dogs that
• 44:40have both have different survival.
• 44:42But we do get better
• 44:44survival again, treating dogs with
• 44:46standard of care, either chemo
• 44:49or surgery and chemo along
• 44:50with EGFR
• 44:52neoantigen therapy.
• 44:56Bladder cancer
• 44:58that glows with,
• 45:00antibodies that arise from EGFR
• 45:02neoantigen therapy. Those on the
• 45:04top versus,
• 45:06non cancer tissue on the
• 45:07bottom control bladder. Actually, it's
• 45:09yeah.
• 45:11Non tumor tissue.
• 45:13So how do you begin
• 45:14to think about this in
• 45:15terms of translating this into
• 45:17human neoantigen therapies. First, you
• 45:19want to know
• 45:20if it potentially this neoantigen
• 45:22that we have been studying
• 45:25in dogs will potentially bind
• 45:27human HLA
• 45:28proteins.
• 45:29That's what you need to
• 45:30get an immune response. This
• 45:32is an algorithm
• 45:34that we put our neoantigen
• 45:36sequence, amino acid sequence through.
• 45:39And about ninety five percent
• 45:41of humans will bind theoretically
• 45:44now. And the algorithms are
• 45:45not perfect.
• 45:46But, it's predicted that most
• 45:49humans,
• 45:50HLA that express these HLA
• 45:53a alleles will bind this
• 45:54neoantigen,
• 45:57as well as class two
• 45:58HLA d q, to a
• 46:00lesser extent, more than half
• 46:02though.
• 46:03So that's really as far
• 46:05as we've gotten in trying
• 46:07to translate this into human
• 46:08disease.
• 46:10We'll see what happens in
• 46:11the future.
• 46:14I don't have to explain
• 46:15to this audience what,
• 46:16checkpoint inhibitor therapies look like.
• 46:19And I won't have time
• 46:20today to go over this
• 46:22data yet.
• 46:24Merck has developed the first
• 46:26checkpoint inhibitor for use in
• 46:28canine cancers. It's an anti
• 46:29PD one.
• 46:31We have an ongoing clinical
• 46:32trial in hemangiosarcoma
• 46:35combining anti PD one with
• 46:37our EGFR neoantigen
• 46:39therapy to see if the
• 46:41combination
• 46:42therapy works better than either
• 46:44therapy alone.
• 46:46We just started this several
• 46:47months ago and we really
• 46:48don't have enough patients yet
• 46:50for statistical
• 46:52significance.
• 46:54So we're not the first
• 46:55to think of this, and
• 46:56you've probably seen lots of
• 46:58other strategies that utilize,
• 47:01neoantigen therapies in humans.
• 47:03And if you look at
• 47:04clinical trials dot gov, there
• 47:06are about a hundred and
• 47:06ninety or so different trials
• 47:09either using personalized neoantigens
• 47:11or DNA
• 47:13or
• 47:14potentially
• 47:15dendritic cell vaccines.
• 47:17Why
• 47:18would one want to use
• 47:19this? Well, at least in
• 47:21our case, cost of production
• 47:22is significantly less than things
• 47:24like monoclonal antibody therapies.
• 47:27It's more easily administered injected,
• 47:30not requiring long term IV
• 47:32therapy.
• 47:33The side effects are minimal.
• 47:35A little swelling at the
• 47:36site, at least in our
• 47:37dog
• 47:40patients. Bunch of happy patients,
• 47:42which
• 47:44I sleep well at night
• 47:45for.
• 47:49So and this is,
• 47:51this is probably the most
• 47:52fun I've had in a
• 47:53long career here at Yale
• 47:55is getting to see patients
• 47:56doing very well dog patients.
• 47:59We have a very unique
• 48:00one in the lower right
• 48:01hand side,
• 48:03which I'll show you in
• 48:04just a minute here. This
• 48:05was a dog that had
• 48:06osteosarcoma,
• 48:08front limb
• 48:09amputation,
• 48:11had metastases
• 48:13to another rear limb,
• 48:15was given our therapy about
• 48:17the same time as the
• 48:19second amputation.
• 48:20This is what the owner
• 48:21chose to do.
• 48:23Let's see if I can
• 48:24get this to work. Behold,
• 48:26the single most perfect thing
• 48:28I have ever seen. This
• 48:30is a dog that's entire
• 48:31life. Still surviving about three
• 48:33and a half years later.
• 48:35So,
• 48:37ranger is right next to
• 48:39that dog. Ranger was another
• 48:41three and a half year
• 48:42survival,
• 48:43had a lung met, front
• 48:44leg amputation,
• 48:46and did very well for
• 48:47a long period of time.
• 48:50Unfortunately,
• 48:51we didn't and we didn't
• 48:52know what Ranger ultimately passed
• 48:54away from. May have been
• 48:56a
• 48:56a rogue metastasis to the
• 48:58spine.
• 49:00It's difficult to tell again
• 49:01in a lot of these
• 49:02dogs. We unfortunately
• 49:04are not able to get
• 49:05certain tissues.
• 49:08So why are some of
• 49:09these things important? Well, our
• 49:11neoantigen eGFR therapy may,
• 49:14just like using
• 49:16monoclonal antibodies,
• 49:18increase the efficacy of other
• 49:20adjuvant therapies like checkpoint inhibitors
• 49:23or radiation
• 49:26that's illustrated in another,
• 49:28another group of studies, not
• 49:30only here, but elsewhere.
• 49:33And our future studies
• 49:35are really to,
• 49:37again, define how this therapy
• 49:39we're not claiming it's gonna
• 49:41be a standalone therapy. Perhaps
• 49:42how it works better with
• 49:43standard of care with radiation
• 49:45or other checkpoint inhibitor therapies.
• 49:48We'd like to know how
• 49:49the tumor microenvironment,
• 49:50just like those of you
• 49:52that study human tumor microenvironments,
• 49:54how that,
• 49:55affects efficacy,
• 49:58of these therapies.
• 49:59And ultimately,
• 50:00understanding individual cell populations
• 50:03in these
• 50:04tumors. A lot of people
• 50:06to thank,
• 50:07a few of them in
• 50:08the audience here, Hester and
• 50:09Ranil.
• 50:10We have twelve different sites.
• 50:13We can't do anything without
• 50:14them. They're scattered around the
• 50:16United States.
• 50:19And with that, I'd be
• 50:20happy to take questions.
• 50:22Yeah.
• 50:26I'll
• 50:29start with the first.
• 50:30What is the
• 50:32issue with inbred drug streams?
• 50:34Imagine they are quite divergent.
• 50:36Does that affect
• 50:37response to it?
• 50:40Question is,
• 50:42are
• 50:43but I I assume you're
• 50:45asking whether tumors are potentially
• 50:47inbred.
• 50:48And the answer is yes.
• 50:50Or the immune system. Yes.
• 50:51Yes to both.
• 50:54We
• 50:55have the data
• 50:57to dive into. So for
• 50:58example, all of our red
• 50:59cap data will illustrate which
• 51:02ones are potentially purebred versus
• 51:04mixed breed,
• 51:05and we can start to
• 51:07follow lineages.
• 51:09There is a golden retriever
• 51:10lifetime study that an organization
• 51:13does that measures
• 51:14frequency
• 51:15of different tumors just in
• 51:17golden retrievers,
• 51:19because it's a popular dog.
• 51:20There's another
• 51:22biobank for Labrador retrievers.
• 51:25So
• 51:26the literally,
• 51:28we're not yet quite sure,
• 51:30but that data is being
• 51:31collected along with the genomics
• 51:33data that the NCI is
• 51:35collecting.
• 51:38How much is the p
• 51:39d one inhibitor? How much
• 51:40does it cost?
• 51:44That is a great question.
• 51:45How much does p d
• 51:46one antibody for dogs cost?
• 51:51Which brings up another topic
• 51:52that differs, of course, between
• 51:55human
• 51:56and canine cancer care, which
• 51:57is economics. Right?
• 52:00A lot of people
• 52:02one flaw in the data,
• 52:03which you've forced me to
• 52:05admit
• 52:07to, is that,
• 52:09again, because treating your dog
• 52:11is driven by economics, they
• 52:12get our therapy for free.
• 52:15We ask them to participate,
• 52:17get surgery, get chemotherapy,
• 52:19and that's an out of
• 52:20pocket expense to them unless
• 52:21they have that cancer,
• 52:23insurance, which very few people
• 52:25only about five percent of
• 52:26dog owners in this country
• 52:28have that insurance.
• 52:30So to your question,
• 52:32it's a lot cheaper than
• 52:33human
• 52:35checkpoint inhibitors, as you can
• 52:37imagine, but it's not cheap.
• 52:39It is based on the
• 52:41size of the dog
• 52:42because it's a MIG per
• 52:44kg,
• 52:45therapy.
• 52:46But anti PD one from
• 52:47Merck will cost anywhere from
• 52:50and there are six or
• 52:52so six or eight, six
• 52:53to ten monthly
• 52:55administrations,
• 52:57of anti p d one.
• 52:59And
• 53:00a total expense, if you
• 53:01have a small dog,
• 53:03six or eight thousand,
• 53:05you've got big dogs, fifteen
• 53:07to twenty thousand for all
• 53:08of the therapy.
• 53:11I don't know if pet
• 53:12insurance even covers that yet
• 53:13even though it's an approved
• 53:15therapy.
• 53:17Patients human patients go to
• 53:19Petco and buy antibiotics.
• 53:21Yep. You got you don't
• 53:22need a prescription,
• 53:23and, you know, they you
• 53:25can just calculate the the
• 53:26human dose for antibiotics.
• 53:28So I know this is
• 53:29a
• 53:30antibody.
• 53:32Yeah, this is different. I
• 53:33understand, but I was just
• 53:34curious about the economics of
• 53:36developing these drugs because I
• 53:37know people are spending a
• 53:38lot more money on their
• 53:39animals too. Yeah.
• 53:42Well, I I didn't mention
• 53:43this. There are two companies
• 53:45out there that do
• 53:47t cell directed basically, CAR
• 53:49T e cells. They expand.
• 53:51They get tumors.
• 53:52I won't mention the competitor
• 53:54companies. They get tumors from
• 53:55the dogs.
• 53:56They
• 53:57extract
• 53:58t cells. They grow them
• 53:59up,
• 54:00send them back
• 54:01to the clinic,
• 54:03and infuse
• 54:04c d eights or c
• 54:05and or c d fours.
• 54:07That costs about,
• 54:09twenty thousand.
• 54:12But there are
• 54:13couple of companies that are
• 54:15trying to make
• 54:16a a go of that.
• 54:18There is another company that
• 54:19will take
• 54:20a tumor tissue,
• 54:23basically, make a gross cell
• 54:24lysate, construct a vaccine, and
• 54:26give that back to the
• 54:27the canine patient as well.
• 54:29So
• 54:30but, unfortunately, the
• 54:33do any anybody here with
• 54:34a dog with cancer? If
• 54:36there are, you know how
• 54:38antiquated
• 54:40therapies are for dogs. They
• 54:42just have lagged far, far
• 54:43behind
• 54:44treatments in humans. Even though
• 54:46a lot of these human
• 54:47drugs are now being used
• 54:50off label and dogs
• 54:51still don't know how they
• 54:52work. Pharmacokinetics
• 54:54are probably very different
• 54:57dosing and etcetera. So,
• 54:59it's
• 55:01they just are not yet
• 55:02good therapies for breeding dog
• 55:04cancers.
• 55:06Any other questions?
• 55:07Yes. Yep. So and you
• 55:09may have said this, and
• 55:10I I missed it, but
• 55:12the selection of your particular
• 55:13EPF R
• 55:15neoantigens,
• 55:16I assume that was from
• 55:17some
• 55:18one of those genomic proteomic
• 55:20screens.
• 55:22But have you looked at
• 55:23a
• 55:26looking at actual new
• 55:28neo mutations as opposed to
• 55:30just particular septide?
• 55:33Obviously, you'd have to then
• 55:34make sure that the cancer
• 55:36that that that's the animal
• 55:38had that mutation.
• 55:39Yeah.
• 55:40But looking at that in
• 55:41versus
• 55:42combinations of neoantigen
• 55:44peptide, etcetera.
• 55:47Yeah. That's a terrific question.
• 55:48Either combination of peptides or
• 55:51looking at high frequency
• 55:54mutations
• 55:54that lead to neoantigens.
• 55:57That can be done.
• 55:59And in fact,
• 56:00I've been using that genomics
• 56:02website
• 56:03to try and define
• 56:05high frequency, for example, osteosarcoma,
• 56:07EGFR based mutations.
• 56:10I think the databanks not
• 56:11yet large enough to narrow
• 56:13it down to defining specific
• 56:15neoantigens.
• 56:17I don't
• 56:18think canine therapy will ever
• 56:19get to the point of
• 56:21personalized
• 56:22neoantigen,
• 56:23mutational neoantigen care.
• 56:26However,
• 56:27I could be very wrong
• 56:28because there are
• 56:31three companies that do canine
• 56:33genomics.
• 56:34So you can ask your
• 56:36vet to send tumor to
• 56:37three different companies.
• 56:38They'll send
• 56:40the owner,
• 56:42mutation analysis.
• 56:43And potentially drugs
• 56:45that may best
• 56:49be effective in that particular
• 56:50dog. Now the drugs may
• 56:51not be
• 56:53available,
• 56:54right, to the vet or
• 56:55to the dog.
• 56:57They're all based on human
• 56:59algorithms for treating a particular
• 57:01mutation.
• 57:02But the companies exist. They're
• 57:04out there.
• 57:06Thank you very much. Thanks.