Mystery Rays from Outer Space

Meddling with things mankind is not meant to understand. Also, pictures of my kids

February 3rd, 2010

Tumors as ecosystems

Park et al JCI 2010 Fig 2
Clonal evolution during in situ to invasive breast carcinoma progression1

What’s a tumor?

In some ways, that’s a bad question (never mind the answer) because it implies that a tumor is a single thing. But we know that’s not true. A tumor, by the time we can detect it, is a collection of many cells, at least billions of them, and those cells are not all the same. I’m not even talking about the normal cell types that are incorporated into a tumor (things like blood vessels and support cells). Even cells that are unambiguously cancerous are very different within a tumor. And of course, that’s important for the things we’re most interested in, prognosis and treatment, because it’s not the average tumor cell that we’re most concerned about, it’s the subset of tumor cells that are most resistant to treatment, or that are most aggressive.

The development of this variation is really fundamental to how we understand tumor formation and tumor growth. Cancerous cells don’t just appear, fully ready to metastasize and grow. What happens is that a normal cell mutates slightly and gains a little advantage. Most of its progeny stay like that, but one of them mutates again and changes a little more, and then one of that cell’s progeny mutates again, and so on. It probably takes at least a half-dozen mutations, over many cell generations, before a normal cell has progressed through to a detectably cancerous cell.  (I’ve talked about this before, here.)

Also, since truly normal cells simply don’t mutate that many times — there are too many checks and repair systems to allow a half-dozen mutations to accumulate in a single human’s2 lifetime — one of the mutations is probably in the check/repair system, turning the cancerous pathway into a mutator pathway as well.

So we expect tumors to be made up of many different cell types, and this is indeed what we see:

With rare exceptions, human malignancies are thought to originate from a single cell, yet by the time of diagnosis, most tumors display startling heterogeneity in cell morphology, proliferation rates, angiogenic and metastatic potential, and expression of cell surface molecules. 1

So how diverse are tumors?

That’s been a hard question to answer, because you’d need tools to look at individual cells, and you’d also need some way of expressing that diversity. A recent paper1 looked at diversity in breast cancer using some individual-cell tools, which I’m not going to discuss, and took an interesting approach to describing the variability:

… we applied diversity measures from the ecology and evolution sciences to our copy number data. These diversity measures estimate the number and distribution of species in a certain geographical area or environmental niche. In our context, a species is a cancer cell population … Hence, a region of a tumor containing cancer cells with 3 different copy number ratios is interpreted to contain 3 distinct “species.” 1

They suggest that this way of describing tumors could be a useful aid to prognosis and to predicting response to therapy, offering a quantitative description of tumor variability (which might correlate with the tumor’s potential for spread and escaping treatment).

I hadn’t thought of tumors as ecosystems before, but I wonder if the analogy could be taken further by considering, say,cytotoxic T lymphocytes as predators …


  1. Park, S., Gönen, M., Kim, H., Michor, F., & Polyak, K. (2010). Cellular and genetic diversity in the progression of in situ human breast carcinomas to an invasive phenotype Journal of Clinical Investigation DOI: 10.1172/JCI40724[][][][]
  2. let alone a mouse’s lifetime[]
November 4th, 2009

Tumor TRegs are more focused than I expected

TRegs infiltrate a tumor
TRegs infiltrate into a tumor

One of the reasons the immune system doesn’t destroy tumors is the presence of regulatory T cells (TRegs) that actively shut down the anti-tumor response.  For once, there’s a little bit of encouraging news on that front.

TRegs are normal parts of the immune system.  They actively prevent other T cells (and so on) from attacking their target. 1  What’s more, TRegs are antigen-specific.  That is, they recognize a specific target, just as do other T cells, but instead of responding by, say, destroying the cells (like  cytotoxic T lymphocyte) or by releasing interferon (like a T helper cell) a TReg’s response to antigen is to prevent other T cells from doing anything in response to that antigen.  In other words, TRegs cause an antigen-specific inhibition of the conventional immune response. 2

Back to tumors.  We know that immune responses don’t routinely eliminate tumors by the time they’re detectable.  There is some evidence that lots of very small, proto-tumors, are in fact destroyed by the immune system very early on, before they’re clinically detectable, but those tumors that survive that attack seem to be pretty resistant to immune control.  At least part of that resistance is because TRegs get co-opted into the tumor’s control (see here, and references therein, for more on that).

So if TRegs are antigen-specific, and TRegs control immune responses to the tumor, what are the tumor antigens that are driving the TRegs?

I would have assumed that TRegs are looking at many, many tumor antigens, including both normal self antigens3 as well as classical tumor antigens.4  But a recent paper5 suggests, to my surprise, that this assumption is wrong.  Instead, “Tregs in tumor patients were highly specific for a distinct set of only a few tumor antigens“. 5 What’s more, eliminating TRegs cranked up the functional immune response, but only to those antigens TRegs recognized — as you’d expect, if the suppression is indeed antigen specific.

This is interesting for several reasons.  If TRegs can be specific for tumor antigens, then at least in theory ((In practice, we don’t quite have the tools yet, I think) it should be possible to turn off these TRegs while leaving the bulk of TRegs intact (and therefore not precipitating violent autoimmunity).  It also suggests that if the TRegs are only suppressing a subset of effector T cells, there’s something else preventing most effector T cells from, well, effecting.  Maybe those are antigen non-specific TRegs, or maybe there’s something else we need to know about.

I’d like to see this sort of study replicated, and a little more fine-tuning on identifying the TReg’s targets (the readout was intentionally fairly coarse here, in order to identify as many as possible).  Still, it’s an unexpected, and potentially very useful, observation.


  1. It’s still not quite clear how they do this[]
  2. There are also antigen-nonspecific TRegs, but we will ignore them for now.  They’re not as effective as the antigen-specific sort, anyway.[]
  3. Because TRegs, unlike most immune cells, can be stimulated by normal self antigens[]
  4. That is, antigens that are mutated, or dysregulated, and that therefore act as standard targets for immune cells[]
  5. Bonertz, A., Weitz, J., Pietsch, D., Rahbari, N., Schlude, C., Ge, Y., Juenger, S., Vlodavsky, I., Khazaie, K., Jaeger, D., Reissfelder, C., Antolovic, D., Aigner, M., Koch, M., & Beckhove, P. (2009). Antigen-specific Tregs control T cell responses against a limited repertoire of tumor antigens in patients with colorectal carcinoma Journal of Clinical Investigation DOI: 10.1172/JCI39608[][]
September 18th, 2009

Beautiful tumors

A new technique called optical frequency domain imaging (OFDI) provides amazing images of tumors (especially their blood vessels) in situ.

3D tumor vasculature with OFDI
“(a) OFDI images of representative control and treated tumors 5 d after initiation of antiangiogenic VEGFR-2. The lymphatic
vascular networks are also presented (blue) for both tumors. (b) Quantification of tumor volume and vascular geometry and
morphology in response to VEGFR-2 blockade.”

Vakoc, B., Lanning, R., Tyrrell, J., Padera, T., Bartlett, L., Stylianopoulos, T., Munn, L., Tearney, G., Fukumura, D., Jain, R., & Bouma, B. (2009). Three-dimensional microscopy of the tumor microenvironment in vivo using optical frequency domain imaging Nature Medicine DOI: 10.1038/nm.1971

August 21st, 2009

Vertical transmission of tumors

Pregnant woman (Ivory Coast)
Pregnant woman (Ivory Coast, West Africa)

Recently I’ve mentioned a few cases of transmissible tumors — that is, cases where tumors actually spread from their original host, to other individuals. The two most dramatic transmissible tumors are Canine Transmissible Venereal Tumor (CTVT) and Tasmanian Devil Facial Tumor (TDFT), where the original tumor can spread widely throughout the entire species. (See this post, this one, and this one, for more detail.) There’s also at least one case of a tumor that accomplished a single transmission, from the original patient to the surgeon who operated on him. 1

Tumors aren’t supposed to be able to spread in this way, because they’re essentially foreign transplants — they should be rapidly rejected, as if they were, say, a skin graft between two random people. In this post I talked about how CVTV and TDFT might have arisen. (There are also a number of cases of tumors that spread to immuno-suppressed individuals, such as after organ transplants, but those cases are easier to understand from an immunologic viewpoint.)

When checking up on references for the last post, I ran across a set of transmissible tumors I hadn’t known about: Vertically transmitted tumors, in which tumors spread from a pregnant mother to the fetus in utero.2

This also, I’m glad to say, seems to be very rare, as you’d expect. Even though mother and child are partially tissue-matched, and even though pregnancy is a very special situation, immunologically, the parent and her child are not genetically identical, and should reject grafts from each other pretty efficiently. (Transplants from parent to child still require immune suppressive treatment.) The review I ran across lists a total of 14 cases of vertical spread of tumors, from 18663 to 2002.4 Although they do note that:

Given the lag time between birth and diagnosis in several of the infants, cases of maternal–fetal transmission may not be as rare as the literature would suggest, and the number of cases could be higher as the detection of metastatic tumor in the fetus may go undetected in cases of abortion or maternal–fetal demise. 2

Malignancy during pregnancy isn’t all that uncommon (0.1% of pregnancies, it says here), so the handful of cases with actual spread of the tumor to the fetus are “numerically inconsequential”. What was different about these 14 cases? We don’t really know, in general. Almost all of the described cases are earlier than 1965,5 predating the molecular era of medicine. Perhaps some, or many, of the infants were immune compromised, as the authors note:

Fetuses with a congenital immunodeficiency are likely to be at an even higher risk for the engraftment of such tumor cells.6 Other factors that may affect the likelihood of tumor cells entering the fetal circulation include maternal homozygosity for one of the fetal HLA haplotypes,7 metastatic potential of the maternal tumor, and a high maternal blood and/or placental tumor load. 2

The outcome of this transmission was very poor; only 3 of the 14 children survived the disease.

I don’t really have any lesson to draw from these cases. Without an extensive molecular workup that isn’t available for almost all of these cases, I don’t know that we can learn much about tumor transmission. Still, these stories are worth keeping in mind when thinking about mechanisms of tumor transmission.


  1. Gartner HV, Seidl Ch, Luckenbach C, et al. Genetic analysis of a sarcoma accidentally transplanted from patient to a surgeon. N Engl J Med 1996;335:1494–1496.[]
  2. Tolar J, & Neglia JP (2003). Transplacental and other routes of cancer transmission between individuals. Journal of pediatric hematology/oncology : official journal of the American Society of Pediatric Hematology/Oncology, 25 (6), 430-4 PMID: 12794519[][][]
  3. Friedreich N. Beitrage zur pathologie des Krebses. Virchows Arch 1866; 36:465–477.[]
  4. Tolar J, Coad JE, Neglia JP. Transplacental transfer of small cell carcinoma of the lung. N Engl J Med 2002; 346:1501–1502.[]
  5. Not saying the molecular medicine abruptly switched on in 1965, it’s just a convenient cutoff[]
  6. Pollack MS, Kirkpatrick D, Kapoor N, et al. Identification by HLA typing of intrauterine-derived maternal T cells in four patients with severe combined immunodeficiency. N Engl J Med 1982; 307:662–666.[]
  7. Osada S, Horibe K, Oiwa K, et al. A case of infantile acute monocytic leukemia caused by vertical transmission of the mother’s leukemic cells. Cancer 1990; 65:1146–1149.[]
August 13th, 2009

Why aren’t most tumors transmissible?

Canine Venereal Tumor phylogeny
Canine Venereal Tumor phylogeny

Bayman commented, after reading this post:

So isn’t the real question why can’t all tumors be transmissible? If you believe the tumor immunologists, all tumors should be capable of avoiding T cell attack…no??

I don’t have answers, but I can speculate a little. 1

Very quick background: In general, tumors are unique. They arise independently each time, and when their host dies, the tumor dies too. That’s in contrast to pathogens, whic may or may not kill their hosts, but which survive and are transmitted to a new host; pathogen infections are not unique, they have a long evolutionary history reaching back through many individual hosts. Tumors can’t do this, for the same reason that skin grafts are rejected by unrelated animals — tumors are essentially unrelated grafts, and should be very rapidly rejected by the new host.

But in very rare circumstances — there are two known instances, and a couple of other possible ones — tumors have arisen that can be transmitted from one host to another.  The two cases are canine transmissible venereal tumor, and Tasmanian Devil facial tumor.  There have been suggestions that these tumors are unique in some immunological way, but I am not convinced by those arguments: See this post and this one for more background.  That’s not to say that these tumors have no ways of evading the immune system; what I am saying, is that virtually all tumors have some way of evading the immune system, and the functions that have been convincingly described for the transmissible tumors don’t seem all that exceptional for tumors in general.

So, if these tumors can be transmitted, and they aren’t all that extraordinary immunologically, what does make them extraordinary?  As I say, I don’t know, but based on tumor immunology as I understand it, I can make some guesses.

The most important factor, I suspect, has nothing to do with immunology.  These tumors are unusual in that they have a built-in way of contacting new hosts. TDFT is spread through bites, CTVT is spread sexually.  There’s no similar way that, say, a liver tumor, or a brain tumor, could be spread.  So that immediately rules out the vast majority of tumors; even if they could survive after transmission, there’s no chance of a transmission chain. 2  But still, most tumors would be rejected even if they did manage to be transmitted.

The Three E's of tumor immunity
The Three E’s of tumor immunity

What seems to happen with most tumors3 is that proto-tumors appear quite early, but are controlled by the immune system – perhaps for years — and never become detectable.  Many such proto-tumors are completely eliminated by the immune system, and we have no way of telling that they even existed.  Many more are controlled at the half-dozen cell stage, much too small to detect; they aren’t eliminated by the immune system, but they can’t escape and grow either.  A very small percentage of these equilibrium tumors, though, eventually find a way of at least partially escaping from immune control, and begin to grow. (Perhaps the immune system kills 99% of the new cells, but a 1.01% growth rate compounds itself fast enough to be eventually detectable.)  This is the “Three E’s” theory of tumor growth (discussed more here and here) — “Elimination, Equilibrium, Escape”.

Regulatory T cells
Regulatory T cells and cancer

The Three E’s apply to the very small proto-tumors. But there is probably another factor that kicks in once the tumor becomes larger.  Tumors are themselves immunosuppressive — they shut down immunity throughout the entire body, to some extent, but they shut down immunity to themselves very powerfully.  The immune system has powerful safeguards that prevent it from attacking its own body; broadly speaking, tumors are their own body, and in many cases tumors probably also have been selected to massively amplify the normal protective signals. 4 (See this post, and this one, for more on that.)

So here5 is my speculation.  We suspect that the ability to be transmitted is present in several tumors, but they never get the opportunity to transmit.  Of those rarities that do get transmitted, most are rapidly rejected, as foreign grafts.  But a tiny minority of this minority may be able to survive because they have powerful immune suppression abilities on top of their common immune evasion abilities.

Tasmanian Devil crossing
Why did the Tasmanian Devil cross the road?

Were CTVT and TDFT just lucky — just happened to have the right immune suppressive abilities?  I don’t think so.  I think they were in the right place at the right time.  They were tumors that had a mechanism for transmission, and that had some ability to immune suppress, but they would normally have been rejected as foreign grafts.  Except that both of these tumors, I think, arose at a time and place where their population was highly inbred.  CTVT arose, we speculate, as dogs were becoming domesticated; probably a small, inbred, closely-related population.  Tasmanian Devils in general may not be closely related, but I suspect there are sub-populations6 that were closely related and that would not have rapidly rejected skin grafts.  The early version of the respective tumors would not have been rapidly rejected by these closely-related new hosts, giving them a chance to establish their own immune suppressive regime.

Now we have the chance for natural selection of the tumors.  Variants with more powerful immune suppression could spread to a wider range of hosts; variants with standard immune suppression died out with their victims.  In dogs, this natural selection could occur over time; as dogs became gradually more variable, there would be continuous new selection for new tumors that could keep up with the dogs. 7 With the Devils, the selection would be over space: The tumors would be selected for their ability to spread within new sub-populations of the Devils, perhaps through gradually more distantly-related subgroups. Eventually, we see the tumors as being capable of transmission and growth throughout the entire population, but the original tumor might not have had this ability.

Channel Island Fox
Rapid MHC diversity in Channel Island Foxes

This model suggests that humans are probably not at great risk of having a transmissible tumor spread in us; and the same is true for most species.  You need the combination of an inbred sub-population with a mechanism of tumor spread and the right kind of tumor. And inbred populations are usually a transient thing; MHC becomes diverse very rapidly, and then the window for tumor establishment is closed.

But this is just a guess, so don’t be too comforted.


  1. And by the way, I disagree with Bayman’s suggestion here (“Tumor Immunology Is A Waste of Time”) that he “find[s] it impossible to believe that effective therapy will ever achieved by artificially stimulating the immune system to attack weak and largely self antigens.” But this post is already too long, so I’ll save my answer for another time.[]
  2. There’s at least one case of a surgeon who apparently contracted a patient’s tumor after cutting himself during surgery — given the option, perhaps many more tumors could be transmissible, but don’t get the chance.[]
  3. Not necessarily those induced artificially, with high doses of carcinogens or with powerful oncogenes, but with those that arise naturally, in older individuals[]
  4. I suspect that tumors have many ways of achieving this localized immune suppression.  I also suspect that different tumors have different dependence on this localized immune suppression.  Those tumors that were highly successful as proto-tumors might already be very good at avoiding immunity — for example, they may secrete tons of TGF?, or otherwise have very powerful TReg-inducing abilities — and only need to shut down a little.  Those that barely squeaked by as proto-tumors, may have very potent immune suppression.  I don’t think the mechanisms for this tumor-based immune suppression are very well understood, though over the next couple years they probably will be. []
  5. Finally![]
  6. Subpopulations that are now, probably, extinct, because of the tumors[]
  7. I’m told that CTVT is eliminated faster or slower in different dogs.  It would be very interesting to correlate this with MHC types, to see if there’s still some effect of rejection even after 50,000 years of selection on these tumors.[]
July 15th, 2009

Transmissible tumors: Similar after all?

Tasmanian Devil road signA year or so ago, the first time I mentioned transmissible tumors, I dismissed Tasmanian Devil Facial Tumor disease as not particularly surprising. As I said more recently, I’ve changed my mind about that, almost entirely because of a chat with Elizabeth Murchison at the Origins of Cancer symposium last month, who told me a couple of things I hadn’t known about the situation.

TDFT is a transmissible tumor, a cancer that arose in one Tasmanian Devil some time ago (at least a decade ago, and maybe much longer) and that is now spreading throughout the Tasmanian Devil population — killing a vast number of them. It’s a tumor that’s spread when these highly territorial animals1 bite each other on the face; the tumor then grows on the bitten animal’s face and eventually makes it unable to eat properly. Nearly 90% of Tasmanian Devils, in affected areas, have died. This is a catastrophe.

(The good news, such as it is, may be that the Devils as a population are adapting to the tumor to some extent — not by resisting the tumor, though, but through rapid selection for much earlier breeding. 2 By reproducing earlier, the population may be maintained even though the Devils are dying as adults. Of course, the population that “survives” won’t be the same as those the existed earlier — they’ll be younger, they’ll have all kinds of different characteristics because they’re no longer being selected for adult survival — but at least it might open a window to save the species.)

Orphan Tasmanian DevilCancers are not, in general, transmissible.  Each new cancer is a new event, that arises from normal cells of the affected individual.  A transmissible cancer would be like an organ transplant — it would be rapidly rejected, because individuals in a population have different MHC molecules, and the immune system rapidly and aggressively rejects cells with the wrong MHC. So there are two possible reasons why a tumor could, in fact, become transmissible. Either the individuals in a population do not have different MHC molecules on their cells, or the tumor can transmit between individual in spite of different MHC molecules.

The explanation for the spread of TDFT seemed to be the former: It was claimed that Tasmanian Devils have very little MHC diversity, so that they can’t reject the tumors3. Even though MHC is normally highly diverse in a population, there are certainly populations — especially small, island, threatened populations, like Tasmanian Devils — that have very limited MHC diversity, so this seemed reasonably plausible. 4 This would be a fairly well-understood mechanism of tumor spread, which is why I said it wasn’t that interesting.

Tasmanian Devil skeletonThe other possibility is that the tumor could spread between individuals in spite of them having different MHC molecules. This is not supposed to happen, according to our understanding of MHC and organ transplants; but in fact, it’s the way the only other extant transmissible tumor (Canine Transmissible Venereal Tumor) spreads.  We have no idea why that happens, which makes it interesting. (A number of ways have been proposed by which CTVT avoids rejection. I won’t go into them in any detail here: none of them, as far as I can see, are unique to CTVT, but rather are very common molecular changes in tumors of all kinds, and yet other tumors are not transmissible; so I don’t consider any of these suggestions to explain the unique feature of CTVT.)

Anyway, as I say, the explanation for TDFT was simple enough: The tumor spread because Tasmanian Devils aren’t genetically diverse. But there are three major arguments, I’ve learned, that say that explanation is wrong.

First: Tasmanian Devils are not, in fact, spectacularly genetically homogenous. 5 They’re not as diverse as you’d like to see, but they’re not completely homogenous. In particular, there are two clear, genetically distinct, populations of Devils in Tasmania, one in the East, and another in the Northwest.

Second: Murchison told me that Tasmanian Devils — even those in the same sub-population — vigorously reject each others’ skin grafts. This is what’s supposed to happen with skin grafts, of course. It implies that the Devils do not, in fact, have the same MHC; and in my opinion it’s a much stronger experiment than those in the original homogenous-MHC paper. 3 If Devils reject skin grafts from each other, then they ought to reject tumors from each other — in other words, even if the tumor can take in one individual, then it should be rejected in another, so the tumor should not spread throughout the population. The skin graft finding hasn’t, as far as I know, been published, but if it holds up, it’s a strong argument against homogenous MHC.

Third: Murchison also told me that the tumor is spreading into Devils in the Northwest. 6 As I said, the Northwestern and Eastern Tasmanian Devils are clearly distinct populations, with different genetic characteristics. 7  If the tumor can spread between them, then the tumor isn’t relying on genetic homogeneity for its transmission.

So the evidence for MHC homogeneity is not good, the evidence that the tumor requires MHC homogeneity is not good, and the only precedent we know of, CTVT, does not require MHC homogeneity for its transmission.

It seems that the two transmissible tumors we know of may be much more similar than I had thought.


  1. Incidentally, when I was looking for images of Tasmanian Devils, I found it interesting that the great majority of images I found using Google Image Search showed an open-mouthed, angry Devil, about to take a chomp out of something.  I was going to comment on that, and then I went to Flickr and did the same search, finding instead thousands of pictures of peaceful, timid, close-mouthed Devils.  I think the popular images, put by Google at the top of the list, as so popular because they reinforce the Devil stereotype, and the Flickr photos are a more accurate reflection of their true nature.[]
  2. Jones, M., Cockburn, A., Hamede, R., Hawkins, C., Hesterman, H., Lachish, S., Mann, D., McCallum, H., & Pemberton, D. (2008). Life-history change in disease-ravaged Tasmanian devil populations Proceedings of the National Academy of Sciences, 105 (29), 10023-10027 DOI: 10.1073/pnas.0711236105[]
  3. Siddle, H. V., Kreiss, A., Eldridge, M. D., Noonan, E., Clarke, C. J., Pyecroft, S., Woods, G. M., and Belov, K. (2007). Transmission of a fatal clonal tumor by biting occurs due to depleted MHC diversity in a threatened carnivorous marsupial. Proc Natl Acad Sci U S A 104:16221-16226[][]
  4. Olivia Judson had a very good summary of the argument in her blog; she also made the important point that MHC diversity, or lack of it, is clearly not the only factor involved in the tumor spread.[]
  5. JONES, M., PAETKAU, D., GEFFEN, E., & MORITZ, C. (2004). Genetic diversity and population structure of Tasmanian devils, the largest marsupial carnivore Molecular Ecology, 13 (8), 2197-2209 DOI: 10.1111/j.1365-294X.2004.02239.x[]
  6. Again, as far as I know this isn’t published yet.[]
  7. The authors of the 2004 paper didn’t specifically look at MHC diversity, though; so it remains possible, if unlikely, that the MHC is relatively homogenous while the rest of the genome is diverse.  This is completely the opposite of the usual situation, so I think it’s not likely.[]
May 26th, 2009

Why (some) similar tumors are similar

Caco2 colon carcinoma cells
Caco2 colon carcinoma cells

One of my long-standing questions now has at least a partial answer, or maybe a pathway toward a partial answer.

My question was, “Why are different tumors the same?” That is, why do tumors of the same type often seem to have similar immunological changes?

Viruses of the same kind (all herpes simplex viruses, say) all avoid immunity in the same way because they all have a common ancestor from which they inherited their immune evasion molecules. But tumors have no common ancestor; each tumor has to grope around and find its own solution to common problems independently. So why should tumors of a particular tissue converge on common solutions? What would make a certain pathway a simple solution for colon carcinomas, but not for bladder tumors? And yet, apparently that’s what happens. Even though many different tumor types become non-immunogenic, tumors of a particular tissue type often reach non-immunogenicity by a similar path. For example:

… distinct molecular events underlie HLA class I loss, depending on the aetiology of the tumours; Lynch syndrome-related cancers presented with mutations in the β2-m molecule, while sporadic microsatellite-unstable tumours mainly showed alterations in the antigen-processing machinery components 1

When I posed this question a couple of months ago, I didn’t have an answer. I said

Part of the answer may be that the particular oncogenes associated with different tumor types lead to particular transcriptional hot-spots, and being a transcriptional hot-spot makes the region a mutational hot-spot as well, but at least as I understand it that’s not enough to account for the trends.

EH, in the comments, suggested a couple more possibilities: Perhaps destruction of a particular pathway is “a nice side benefit of destroying some yet undiscovered tumor suppressor important for melanoma or colon cancer? Or maybe the loss of certain repair genes common to a cancer type (like BRCA) leads to selective chromosomal instability in parts of chromosome 6?

It turns out, unsurprisingly, that I’m not the only one to ask the question. There is no general answer, but apparently a partial answer has been kicking around for a while now, and Hans Morreau’s group is starting to put some of the pieces together.2 As their model, Morreau’s group looked at MUTYH-associated polyposis (“MAP”)-associated tumors. (MAP is a heritable defect in DNA repair, so MAP patients have high rates of mutagenesis and usually develop colon carcinomas.) They reasoned that

MAP tumours could be more prone to stimulate a cytotoxic T-cell-mediated immune response, due to their frequent generation of aberrant peptides. Hence, these tumours could also be subjected to a strong selective pressure favouring the outgrowth of cancer cells that acquire an immune evasive phenotype.

DNA Repair
DNA Repair

In other words, the argument is that the group of tumors that lose (or reduce) DNA repair are much more likely to throw out mutant proteins. These mutant proteins are targets for the immune response (because they’re no longer “self” antigens), so to survive the immune attack the tumor has a strong selection for loss of immunogenicity (and also has the high mutation rate that allows them to rapidly mutate away from immunogenicity).

Sure enough, the tumors did frequently (72%) have defects in antigen presentation. (In fact, because of the way they measured HLA expression — by immunohistochemistry rather than sequencing — I would bet that the rate of functional defects was actually much higher than that.) They conclude that this “provides additional evidence that tumours carrying defects in DNA base repair mechanisms are more prone to undergo immune escape mechanisms.2  Since they don’t formally compare to other tumor types, I don’t think they can really say “more prone” — you’d have to use the same techniques to look at tumors from non-MAP patients to be able to say that. Still, I do think that is a significantly higher rate than has been turned up in previous studies using similar techniques,3 so I’ll tentatively accept that conclusion.

This doesn’t really explain why similar tumors target similar components, but it’s at least a conceptual connection between different tumor types and an underlying pathway. I’d be interested in a more large-scale screen, looking at cancers with stronger and weaker mutator phenotypes to see if common pathways emerge. One may already have popped up, since the authors note here that expression of β2-m was frequently lost4 in several tumors with DNA repair defects:

Although speculative, it is interesting to underline that carcinomas derived from both MAP and Lynch syndromes preferentially lose β2-m expression coupled to HLA class I deficiencies. A functional explanation for these observations remains elusive, but perhaps distinct reactions (both qualitative and quantitative) by the immune system, depending on the age of onset of the tumours, could condition the type of mechanisms that lead to HLA class I expression deficiencies. 2

Again, we would need to compare to other tumor types to see if this really is more frequent, but overall it feels as if there’s a hint of a pathway here. At least there are some specific questions that can be asked.


  1. de Miranda, N., Nielsen, M., Pereira, D., van Puijenbroek, M., Vasen, H., Hes, F., van Wezel, T., & Morreau, H. (2009). MUTYH-associated polyposis carcinomas frequently lose HLA class I expression-a common event amongst DNA-repair-deficient colorectal cancers The Journal of Pathology DOI: 10.1002/path.2569
    Referencing
    Dierssen JWF, de Miranda NFCC, Ferrone S, van Puijenbroek M, Cornelisse CJ, Fleuren GJ, et al. HNPCC versus sporadic microsatellite-unstable colon cancers follow different routes toward loss of HLA class I expression. BMC Cancer 2007; 7: 33[]
  2. de Miranda, N., Nielsen, M., Pereira, D., van Puijenbroek, M., Vasen, H., Hes, F., van Wezel, T., & Morreau, H. (2009). MUTYH-associated polyposis carcinomas frequently lose HLA class I expression-a common event amongst DNA-repair-deficient colorectal cancers The Journal of Pathology DOI: 10.1002/path.2569[][][]
  3. Other studies, especially those of Soldano Ferrone, have turned up much higher rates of functional HLA class I defects, but they’ve used more focused, and more difficult and expensive, techniques to reach that conclusion.[]
  4. β2-m is a physical component of the MHC class I complex[]
April 14th, 2009

Tumor immunity: The Goldilocks approach

GoldilocksWe know that the immune system can destroy tumors. We also know, unfortunately, that by the time we see a tumor, immunity probably won’t destroy the tumor. There are lots of reasons for that. One is that tumors are essentially part of the normal body, so it’s normal for the immune system to ignore them. It looks as if you need to have immunity that’s just right to get rid of a tumor.

Tumors arise from normal self cells,1 that the immune response has been programmed to ignore. Now, the process of becoming a tumor is not normal, and so tumors are not entirely normal self any more — meaning that there are likely to be some targets in most if not all tumors. But in all but the most reckless tumors the differences between abnormal and normal are relatively small, compared to, say, a virus-infected cell that contains many potential targets.

There’s actually a long list of known tumor antigens; the T-cell tumor peptide database lists many hundreds of them. But most are not truly specific for the tumor.  The’re actually normal self antigens; they’re derived from proteins that are overexpressed in tumors, or that are differentiation antigens or “cancer-germline” antigens that are normally also found in self tissues. What’s more, these normal self antigens are the most interesting tumor antigens, as far as clinical utility is concerned. Mutations can make brand-new, non-self targets for the immune system, but they’re going to be sporadic targets, often unique to individual tumors — not something you can prepare for. The normal antigens, though, are likely to be predictable, common targets; it’s conceivable that tumor vaccines can be prepared in advance.

Melanoma cell (Eva-Maria Schnäker, University of Münster)
Human melanoma cell

If these antigens were common (which they are, in some tumor types — like melanoma), and they were good targets for the immune system, then we wouldn’t see much cancer. We do see melanomas quite often, and part of the reason may be that the immune system generally responds quite weakly to these antigens.  Why is that? And, more to the point, how can we make the immune system respond more strongly? A recent paper in the Journal of Experimental Medicine2 offers answers for both of these questions.

From work in the past couple of years, we now have decent estimates of how many T cells there are that can react with any particular target. (See here and here for my discussion of the earlier papers.) A reasonably strong immune response to a non-self epitope might originate from maybe 100 or so precursor T cells. There’s a rather wide range of frequency for these precursor cells, say from 20 to 1000; and to some extent, the fewer T cells there are the weaker (the less immunodominant) the immune response.

We expect T cells against normal self targets to be less common, because they should be eliminated as they mature in the thymus. Some may survive, though, and we would count on these survivors to attack the normal (albeit overexpressed, or abnormally present) target in the cancer cells. But just how rare are they?

Rizzuto et al say they’re really rare (this was in mice, by the way); at least ten times less abundant than T cells against non-self antigens.  If you look at the range I gave for “normal” precursors, that could mean there are fewer than 5 or 10 precursors.  If the average is “fewer than five”, then quite possibly some mice have only two, or one, or no precursors.  You can’t have much of a response with no precursors.

So there’s a weak anti-tumor response because there aren’t many T cells in the body that can respond to the normal self targets in the tumor. That’s not really a surprise, but it does raise the question, What if there were more of the T cells? To ask that question, Rizzuto et al. tried transferring more of these precursor T cells into tumor-bearing mice — starting at around the normal level for a precursor to non-self antigen, and going up from there — and then vaccinating with the appropriate target.

The effects were pretty dramatic. With no supplemental T cells (that is, with the natural, very low, level of T cell precursors) the mice all died of the tumor quickly. At the middle of the range, almost all of the mice rejected the tumor. And at the highest levels of transfers? The mice all died again. Having enough T cells to respond was protective, but putting in too many made them useless.

These results identify vaccine-specific CD8+ precursor frequency as a remarkably significant predictor of treatment and side-effect outcome. Paradoxically, above a certain threshold there is an inverse relationship between pmel-1 clonal frequency and vaccine-induced tumor rejection.2

Melanoma cell
Mouse melanoma cell

(My emphasis) This paradoxical effect is probably because the numerous T cells started to compete with each other so that none of them were properly activated; they only saw effective-looking polyfunctional T cells at the lower transfer levels.

In other words, if you’re going to transfer T cells to try to eliminate a tumor, more is not necessarily better. Quality and quantity are both important factors, and quantity helps determine quality.

One question I have is how this relates to tumor immune evasion. Many tumor types  acquire mutations, as they develop, that block presentation of antigen to T cells. Are these mutations perhaps only partially effective — giving the tumors sufficient protection against the tiny handful of natural precursors they “expect” to deal with, but not against a larger attack after, say, vaccination — or are they more complete, and protective even if the optimal number of T cells are transfered? I’d guess that it would depend on the tumor, but it looks as if it might be a relevant question and it would be nice to have more than a guess.

Our results show that combining lymphodepletion with physiologically relevant numbers of naive tumor-specific CD8+ cells and in vivo administration of an effective vaccine generates a high-quality, antitumor response in mice. This approach requires strikingly low numbers of naive tumor-specific cells, making it a new and truly potent treatment strategy.   2


  1. I’m ignoring here crazy things like the contagious tumors of Tasmanian Devils and dogs[]
  2. Rizzuto, G., Merghoub, T., Hirschhorn-Cymerman, D., Liu, C., Lesokhin, A., Sahawneh, D., Zhong, H., Panageas, K., Perales, M., Altan-Bonnet, G., Wolchok, J., & Houghton, A. (2009). Self-antigen-specific CD8+ T cell precursor frequency determines the quality of the antitumor immune response Journal of Experimental Medicine, 206 (4), 849-866 DOI: 10.1084/jem.20081382[][][]
April 9th, 2009

Why are different tumors the same?

Hierarchical clustering of breast carcinomas, Turashvili et al 2007
Hierarchical clustering of breast carcinomas1

Something that’s puzzled me for years is why the same kinds of tumors tend to have the same kinds of immune evasion mechanisms. And I’m not going to give an answer, just trying to share the confusion a little.

What I mean is this:

It has been demonstrated that human tumors of distinct histology express low or downregulated MHC class I surface antigens … The distinct frequency of MHC class I abnormalities is caused by total HLA class I antigen loss, HLA class I down-regulation as well as loss or down-regulation of HLA class I allo-specificities. However, the frequency and mode of these defects significantly varied between the types of tumors analysed and could be associated in some cases with microsatellite instability. 2

(My emphasis) As I’ve noted here several times (most specifically here) tumors very often evade the immune system as they mature. Cytotoxic T lymphocytes (CTL) can control tumors in the tumors’ eary stages, but by the time we detect a tumor clinically the tumor is almost always resistant to the immune system. They do this in various ways, including inducing regulatory T cells, but also by mutating themselves to make themselves invisible to CTL (and other components of the immune system, but let’s keep it simpler for the moment).

There are a myriad ways for a tumor to become invisible, at the molecular level.  The MHC class I antigen presentation pathway is long and complex, and for any partiuclar tumor there are likely to be many different bottlenecks, points of attack.  Since tumors are all independent events3, so at first, and even second, glance, there’s no obvious reason why tumors of the same type should find a similar approach.  That is, just because two colon carcinomas look the same histologically in two different individuals, there’s no link between them.  4 Why should colon carcinomas avoid CTL using one set of mutations, while, say, breast cancers use a different set of mutations? Yet apparently, that’s what tends to happen; for example:

Mutations or deletions in β2-m were detected in colon carcinoma (21%), melanoma (15%) and other tumors (<5%). So far, no mutations in β2-m have been found in RCC lesions, bladder and laryngeal tumors despite MHC class I loss or downregulation. … haplotype loss was found in head and neck squamous cell carcinoma (HNSCC) with a frequency of 36%, whereas in renal cell carcinoma (RCC) LOH only occurs in approximately 12% of tumor lesions analyzed. 2

If we saw these patterns only with virus-associated cancers, such as cervical carcinomas and even hepatic carcinomas, there would at least be a common link, but these tumors are not (as far as we know) caused by viruses in humans.

Part of the answer may be that the particular oncogenes associated with different tumor types lead to particular transcriptional hot-spots, and being a transcriptional hot-spot makes the region a mutational hot-spot as well, but at least as I understand it that’s not enough to account for the trends.

So why are particular MHC abnormalities linked to tumor type?  Anyone?


  1. Turashvili et al. BMC Cancer 2007 7:55   doi:10.1186/1471-2407-7-55[]
  2. Seliger, B. (2008). Molecular mechanisms of MHC class I abnormalities and APM components in human tumors Cancer Immunology, Immunotherapy, 57 (11), 1719-1726 DOI: 10.1007/s00262-008-0515-4[][]
  3. barring such weird things as canine transmissible venereal tumor and Tasmanian Devil facial tumors; see here for more on those[]
  4. The comparison is, of course, viruses.  A herpesvirus of chickens, and one of humans, may both use immune evasion mechanisms, but they have a common ancestor even if it’s a couple of hundred million years ago.[]
February 24th, 2009

More chemotherapy and tumor immunity

U-118 glioma cells (Nikon Microscopy Gallery)
U-118 Glioma cells 

In one of the earliest posts I made to Mystery Rays,  I commented on an exciting anti-tumor finding. 1  Basically, the suggestion was that when chemotherapy of tumors works, it doesn’t actually work by killing all the tumor cells; or at least not directly.  Instead, the authors said, chemo works because the dying tumor cells release adjuvants (immune stimulants),2 and what actually eliminates the tumor is the specific immune response to the tumor, driven and enhanced by the immune stimulants.   In other words, the chemotherapy is actually acting like the adjuvant in an anti-cancer vaccine. 

Presumably the reason this form of adjuvant is effective, whereas just giving an adjuvant to a cancer patient is not, is that it’s so tightly linked to the tumor you want the immune system to target.   

In the subsequent year and a half there have been a handful of papers looking more at this phenomenon (not counting the review papers, which have churned out at a great rate). For example, it’s been shown that dying tumor cells are cross-presented efficiently due to HMGB1 and similar immune stimulators.3    Now, there’s a more specific support of the concept. 4 

I don’t know if the Apetoh et al paper was the initial impetus for the latest one, or (more likely) if it was already in progress, but a any rate, unless I’ve missed one, this is the second study that directly looks at HMGB1 effects on tumor regression following tumor cell death, and it reaches basically the same conclusion as the first paper.  This was strictly a mouse study, but they used a highly aggressive brain tumor (glioblastoma multiforme).  As well as chemotherapy they used a gene therapy approach, thus managing to hit four of my Mystery Rays buzzwords (tumor immunity, immunity to viruses, innate adjuvants, and if you’re generous oncolytic viruses as well) all at once.  (What would a Mystery Rays Bingo Card look like?)

Glioblastoma
Glioblastoma

If the model is correct, then what you really want to do when treating tumors is a double whammy — kill tumor cells, and simultaneously attract in immune cells.  Curtin et al did this by injecting two recombinant adenoviruses directly into the tumor; one rendered the infected cells sensitive to chemotherapy, and the other attracted dendritic cells (DC are important cells for initiating immune responses). Following chemo, the tumors regressed dramatically (about half the mice survived, compared to 100% death without treatment).  And both of the viruses were necessary; without the immune system, killing the cells wasn’t enough, and without tumor cell death, the immune system couldn’t do enough. 

The same treatment in mice without TLR2 did nothing.  TLR2 is an innate immune recognition molecule that triggers inflammation in the presence of (among other things) HMGB1, which is released from dying cells.  This is similar to the Apetoh et al paper, and Curtin et al took the studies a little further by showing that TLR2 had to be on dendritic cells (as opposed to the tumor itself). What’s more, when all the parts were in place (tumor cell death, DC  present, DC expressing TLR2) there was a strong specific anti-tumor cytotoxic T lymphocyte response, while without the TLR2 stimulation there was little such response.

In other words, they’ve firmed up the probable pathway by which chemotherapy eliminated these tumors. The chemotherapy killed cells directly, and the dying cells released HMGB1; the HMGB1 activated dendritic cells in the tumor, by triggering their TLR2 receptor; the activated DC then in turn activated CTL specific for the tumor; and the CTL then completely eliminated the tumor and prevented the remains from regrowing.  (Some of the steps in this pathway remain formally unproven, but the data are consistent with this model.) 

It’s not clear, yet, if this is the universal explanation for successful chemotherapy, or whether chemo sometimes or often works by killing the tumor directly with no requirement for immunity. The authors here looked at several different tumors, expanding on the more correlative data from Apetoh et al., and so far all the cancers that have been checked fall into the immune clearance category.  So, while it’s a long, long way from the bedside, it’s certainly encouraging. 

In conclusion, the results reported provide compelling evidence for the role played by HMGB1 in mediating the efficacy of antiglioma therapeutic regimes that are based on tumor cell killing strategies … [C]ancer immunotherapies coupled with effective cell killing modalities may be necessary to achieve therapeutically relevant antitumor efficacy. 4


  1. Toll-like receptor 4-dependent contribution of the immune system to anticancer chemotherapy and radiotherapy. Apetoh et al. Nature Medicine 13, 1050 – 1059 (2007)  doi:10.1038/nm1622

    Also this review:

    Apetoh, L., F. Ghiringhelli, A. Tesniere, A. Criollo, C. Ortiz, R. Lidereau, C. Mariette, N. Chaput, J. P. Mira, S. Delaloge, F. Andre, T. Tursz, G. Kroemer, and L. Zitvogel. 2007. The interaction between HMGB1 and TLR4 dictates the outcome of anticancer chemotherapy and radiotherapy. Immunol. Rev. 220:47-59.

    and this one:

    Apetoh, L., A. Tesniere, F. Ghiringhelli, G. Kroemer, and L. Zitvogel. 2008. Molecular interactions between dying tumor cells and the innate immune system determine the efficacy of conventional anticancer therapies. Cancer Res. 68:4026-4030.

    and this review from a different group:

    Campana, L., L. Bosurgi, and P. Rovere-Querini. 2008. HMGB1: a two-headed signal regulating tumor progression and immunity. Curr. Opin. Immunol. 20:518-523.[]

  2. Specifically, the dying cells release a protein called HMGB1, which has been shown to be an endogenous adjuvant[]
  3. Brusa, D., S. Garetto, G. Chiorino, M. Scatolini, E. Migliore, G. Camussi, and L. Matera. 2008. Post-apoptotic tumors are more palatable to dendritic cells and enhance their antigen cross-presentation activity. Vaccine 26:6422-6432.[]
  4. James F. Curtin, Naiyou Liu, Marianela Candolfi, Weidong Xiong, Hikmat Assi, Kader Yagiz, Matthew R. Edwards, Kathrin S. Michelsen, Kurt M. Kroeger, Chunyan Liu, A. K. M. Ghulam Muhammad, Mary C. Clark, Moshe Arditi, Begonya Comin-Anduix, Antoni Ribas, Pedro R. Lowenstein, Maria G. Castro (2009). HMGB1 Mediates Endogenous TLR2 Activation and Brain Tumor Regression PLoS Medicine, 6 (1) DOI: 10.1371/journal.pmed.1000010[][]