Each is unique: Intratumor heterogeneity


In the last post, we considered how cancer is more than just one disease.  We learned, for example, that kidney cancer does not necessarily carry the same mutations as breast cancer.  And yet, despite these variations, certain genes were mutated across several cancer types and occurred early in the progression of the disease.  We called these mutations driver mutations.  In this week’s paper this idea will be reiterated in the context of a new concept: intratumor heterogeneity: that each tumor and regions within the same tumor are genetically distinct.  The article that prompts this discussion was published in the New England Journal of Medicine in 2012 (http://www.nejm.org/doi/full/10.1056/NEJMoa1113205).

First: a brief primer on intratumor heterogeneity.   Over time, cells acquire genetic mutations through division errors or external sources that allow them to divide uncontrollably and operate unchecked.  When these cells start to accumulate, we detect a mass or tumor.  Intratumor heterogeneity stems from the hypothesis that each cancer cell has the capability to acquire different mutations due to its own genetic instability.  As these cells expand and migrate, different tumor cell regions with distinct genetic profiles exist.  Additionally, cells that metastasize or migrate to different regions of the body can also evolve into different genetic subpopulations.  This type of progression is also referred to as clonal evolution. 


To assess if this hypothesis holds true, the authors analyzed four patients with renal cell carcinoma (kidney cancer), taking biopsies from multiple tumor and metastatic regions.  By sequencing the coding region of the tumor’s DNA and analyzing the chromosomal copy number, they could identify genetic mutations and abnormal cell duplication, respectively.  For simplicity, we will only discuss Patient 1. 

In addition to the renal carcinoma, Patient 1 had metastases around the kidney area (perinephric), in the lungs and in the chest-wall.  From these areas, the investigators took 15 biopsies including 9 different regions from the primary tumor (See figure). 

                                                               Figure 1 (Gerlinger et al, NEJM 2012)

After sequencing the tumor material, validating the mutations, and checking for false positives and/or negatives, the investigators identified 128 mutations.  Of these, 40 were ubiquitous across every region (only 31%), 59 mutations were shared between most but not all regions, and 29 mutations, called private mutations, were present in only one specific region.  Excluding the metastatic sites and looking within the tumor alone, only slightly more mutations were identified as ubiquitously occurring (34%).  The conclusion? Each region differs slightly at the genetic level.
In addition to identifying mutations, they analyzed the genetic instability of these tumor regions.  One way to do this is by assessing the chromosomal copy number.  Normal cells possess two copies and any aberration from this is called aneuploidy and is a mechanism through which cells can acquire additional mutations.  Ploidy profiling showed that no region shared the same profile, suggesting that every region – possibly every cell – has the capability to change and adapt over time and space.  The authors referred to this concept as clonal evolution and suggest that tumors are continually changing, continually evolving.  Like the theory of human evolution, it is the survival of the fittest.  From these results, it becomes clear:  intratumor heterogeneity is alive and well.
Although this seems like a dire situation, remember that about 30% of regions share the same mutations.  Furthermore, although the type of mutation was different in different biopsies, several genes were commonly mutated across several regions.  For example, in Patient 1, these genes were SETD2 and PTEN.  They also verified that these genes were functionally altered showing that their function is important for maintaining homeostasis.  Finally, the investigators were able to devise a timeline called a phylogenetic tree to depict where and when some of these mutations occurred.  Mutations in specific genes, including VHL, MTOR, and SETD2 could be considered driver mutations.  And it is these genes that we need to focus on. 


Where do we go from here:
I might call intratumor heterogeneity the silent partner in cancer research.  We acknowledge its existence but we have yet to figure out what to do about it.  Moreover, this silent partner only points to its parent: the complexity of cancer. 

As a researcher this means more focus on identifying and targeting driver genes.  According to this publication, 30% of mutations are shared. Are these 30 mutations shared across patients? Moreover, does the clonal evolution of tumors follow a pattern?  For example, does a mutation in gene A lead to a mutation in gene B?  This question probes our ability to predict gene mutations based on existing mutations.   If we can do this, we can design target therapies and combination therapies that may hit multiple evolutionarily tumor branches and better fight the disease.

As a patient this study begs for patience: patience at the clinic where multiple tumor biopsies need to be obtained over multiple time points.  Patience with our governments as we lobby for more money for these added procedures.  And patience with your physician and the behind-the-scenes researchers who penetrate into the intricacies of your tumor development, tailoring your treatment.

Today’s uncovered cancer morsel: Just as each of us is unique, so the tumors that invade our bodies also exhibit singularity.

References:
Gerlinger M., Rowan A., Horswell S, et al. Intratumor Heterogeneity and Branched Evolution Revealed by Multiregion Sequencing.  NEJM.  2012;366(10):883-892.

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