Brisk trade in tissue for proteomics and genomics research
The rapid evolution of clinical genomics has
banking from a relatively informal resource for academic researchers
to a commercial linchpin of the drug and diagnostics industries.
According to industry estimates, the current market for microarray
technology and associated human tissue and RNA is $400 to $800 million—and
on track to double in size soon. Alan D. Proia, MD, PhD, associate
professor and vice chairman of pathology at Duke University, Durham,
NC, noticed the trend a few years back. "I’d get a call a week from
pharmaceutical companies, diagnostic companies, people at Duke and
at Research Triangle Park," he says. "Everybody and his brother
wants human tissue for genomic and proteomic research."
"This is a relatively new branch of tissue banking that’s come
into being in the last five years," says Michael J. Becich, MD,
PhD, chairman of pathology at the University of Pittsburgh Medical
Center Shadyside Hospital and director of the university’s Center
for Pathology Informatics. Dr. Becich, who is also founder of TissueInformatics,
in Pittsburgh, and a former co-chair of the CAP Informatics Committee,
says the number of high-volume formalized tissue banking efforts
around the country has gone from "a handful" 10 years ago to thousands
In 1991, he helped launch what was then called the Western Pennsylvania
Genito-Urinary Tissue Bank, which was housed at the University of
Pittsburgh and drew tissues from a large cluster of hospitals. The
university is now one of four sites funded by the National Cancer
Institute (along with George Washington University in Washington,
DC, the Medical College of Wisconsin in Milwaukee, and New York
University) to maintain national tissue banks for prostate tumor
samples as the Cooperative Prostate Cancer Tissue Resource.
These sites, says Dr. Becich, make more than 3,000 prostate samples
available to researchers, along with serum, whole blood, lymphocytes,
and a rich set of clinical data. And demand for the tissues is surging.
The reason, says Jim Wittliff, MD, PhD, professor of biochemistry
and molecular biology and research professor of surgery at the University
of Louisville (Ky.) James Graham Brown Cancer Center, is, "You now
need human tissue to mine the data that have evolved from the Human
The term "tissue banking" often creates confusion because it is
commonly used to refer both to tissues collected for research and
those used for transplantation. Members of the American Association
of Tissue Banks, for example, are the relatives of organ donation
centers; they meet an equally growing demand for skin, veins, bone,
fetal tissue, and other tissues used in burn treatment, other therapies,
and plastic surgery.
Research-oriented tissue banks, however, do not have to meet standards
for human transplantation and do not have a formal regulatory body.
While collection and preservation standards are essential, these
tissue banks are mainly concerned with the quality of their samples
as scientific information.
Clinical genomics is the correlation of molecular changes—including
differences in patterns of gene expression—with the characteristics
of human disease to accelerate the discovery and validation of targets
for new diagnostics and therapeutic agents. "For the first time
it is moving beyond the thought phase," says Alan Buckler, PhD,
chief science officer of Ardais Corp., Lexington, Mass. "We’re now
marrying genomics and proteomics with actual human disease, and
some very compelling patterns of disease are already emerging in
"Back in 1991," says Dr. Becich, "a lot of people would see the
utility of frozen tissue, but there were a number of problems. To
really create a lot of value for it, you need to provide a rich
clinical annotation, with a lot of information about the patient,
the disease, the organ site the tissue is taken from, and so on."
That required "a lot of heavy lifting," he says, to work through
patient consent issues amid changing regulations to ensure privacy
The new demand for frozen tissue to perform genomic and proteomic
profiling for diagnostic biomarkers and to develop therapeutic targets
has injected the cash to fulfill that task. Relatively new companies
like Ardais, TissueInformatics, and Asterand (in Detroit) cater
to large pharmaceutical and biotech customers—the Lillys,
Abbotts, Pfizers, and Millenium Pharmaceuticals, Dr. Becich says.
And academic institutions are reaping many of the benefits.
At the first Clinical Genomics Symposium, in June 2002, Mark Boguski,
MD, PhD, a computational biologist at Seattle’s Fred Hutchinson
Cancer Research Center, said, "What we are witnessing today is the
shifting upstream towards discovery research of the clinical trial
discipline: namely, the power of large numbers of patient cases
and the systematic application of relevant clinical data to correlate
with molecular change."
Ardais announced in February that AstraZeneca, one of the five
largest pharmaceutical companies in the world, has licensed access
to Ardais’ library of tissue samples and related information for
its drug discovery program. Ardais has commercial agreements with
some 25 other pharmaceutical and biotech companies as well, including
Bristol-Myers Squibb, Aventis, and CuraGen Corp.
It was Oxford Bioscience Partners,
a venture capital fund with
about $600 million invested in 80 biotech companies, that saw the
need for a company like Asterand. "The senior partners, Dr. Alan
Walton and Jonathan Fleming, realized with the mapping of the human
genome that there eventually would be a massive demand for researchers
to actually go beyond the animal models, using mice and so on, that
drug companies used for years as the basis of early research work,"
says Asterand chief executive officer Randal Charlton. "With the
knowledge of how humans are made up, it makes sense to look at gene
and protein expression in the human cells, for which, of course,
human tissue is required. However, scientists in the Oxford group
of companies were not able to find the tissues they needed for this
new approach, and they said, ’Look, we need a special company that
specializes in this discrete task.’"
"Ten years ago," says Dr. Becich, "the number of new biomarkers
in the pipeline as potential new therapeutic avenues probably would
number in the low teens or 20s. Today there are probably 300 to
500 candidates in various stages, making their way through various
frameworks. The majority really come from discoveries with genomic
and proteomic methodologies on tumor tissues."
Adds Charlton: "With the mapping of the human genome, there was
awareness that the growing genomics and proteomics industry would
require well-characterized tissue. In other words, tissue collected
in such a way that genetic information can be extracted. So it had
to be collected under a very strict protocol which we developed
in the first year of our existence. We probably went through 20
iterations of collection protocols." The result was a process in
which the tissue is frozen quickly.
"If it’s diabetes, we’d collect a variety of tissues, including
different fats and samples of other organs, like the pancreas, because
diabetes is not just in one place—whereas for arthritis, we’d
collect synovial fluid," says Charlton. In cancer, he points out,
"the tissues tend to be from the tumor and the adjacent normal tissue,
so there might be several tissues from the same patient."
Just as important as the tissue, he says, "are the clinical information
and pathology information that go with it, and this is what I mean
by ’well-characterized.’ We’ve got complete patient data, both the
patient history and the family history, as well as the pathologist’s
report on the tissue that has been collected."
There are two primary sources for research tissue, Charlton adds.
"One is surgical collection, where someone is having an operation
for colon cancer or lung cancer, for example, and we get surplus
material, in excess of what is required for patient care, that would
otherwise be destroyed. The other source is postmortem or so-called
rapid recovery. What happens is, a patient has preconsented to donate
their body to medical research and our pathology team goes in and
collects material that is required." This collection must be done
rapidly, within 10 hours of death, he notes, and ideally within
six hours or less. Biopsies are another source of tissue, but they
tend not to produce enough tissue to make each sample commercially
After a molecular biologist takes a tiny piece of the sample and
checks its RNA, about eight percent of Asterand’s tissues are rejected
because of degradation—but that percentage rises to 20 or
even 40 percent with postmortem tissue. Surgical material "comes
straight from the operating room to pathology and it’s frozen very
quickly, within minutes," Charlton explains, in contrast to the
several hours allowed for salvaging postmortem tissues.
Founded only three years ago, Asterand receives tissues from 20
sites all over the world and already has about 50 research clients,
the majority of them large pharmaceutical and biotech companies.
The company has 22,000 tissue samples, including all cancer material
(breast, lung, colon, esophageal, prostate, and pancreatic, and
rare cancers) and non-oncology disease targets such as cardiovascular
disease, rheumatoid arthritis, diabetes, Alzheimer’s disease, Parkinson’s
disease, bipolar disorder, schizophrenia, and amyotrophic lateral
"We are very sharply focused as a service company," says Charlton.
"We do not do research ourselves, other than research our customers
ask us to do. We’re different from your normal academic tissue bank,
which basically takes a bunch of material in and hordes it, to put
it crudely. So, for example, we’re not involved in collecting diabetes
tissue and saying we’re not going to sell it because we’re going
to identify the genes that cause diabetes. Some of our competitors
do; we don’t.
"We simply accrue the tissue," Charlton adds, "and it can be frozen
or fixed, or we’ll make it into tissue microarrays, or cut slides
for you, or if you want, we’ll extract RNA and deliver that. We’re
increasingly doing some of that front-end work." A client wanting
to do a study of renal cell carcinoma, for example, may want only
the results from extracting RNA. The company then provides the customer
with digital images of the tissue.
Finding sources for tissue remains a challenge, but companies
frequently rely on established relationships with hospitals to obtain
tissues. "It’s extremely difficult," Charlton says. "Very often
the hospitals themselves want to do their own research, and what
we do is make arrangements whereby in return for their supplying
tissue and becoming part of our donor network, they can have access
to our tissues for their own research. For example, one hospital
we deal with is studying colon cancer, but that’s only 10 percent
of what they’re collecting, so they give us all the other stuff
they do; then they have the right to the rest of our colon cancers."
"We also have a few academic customers who say, ’We can’t afford
to pay the value of the tissue, but if you’ll supply them at a serious
discount, we’ll give you the option on any intellectual property
that results,’" says Charlton.
Ardais Corp., which has more than140,000
samples in its repository,
chose a different strategy. It formed "strategic alliances" with
several established medical institutions by launching the National
Clinical Genomics Initiative, which now includes the medical centers
at the University of Chicago, Beth Israel Deaconess in Boston, Maine
Medical Center in Portland (a teaching hospital of the University
of Vermont College of Medicine), and Duke University.
In a presentation to other potential partners, Ardais said the
advantages of participating in the initiative include access to
Ardais’ research-quality samples and clinical data for academic
research as well as access to a network for commercial genomic research
"For most tissue banking efforts," Ardais’ Dr. Buckler says, "normal
tissue is typically remnant tissue from surgical procedures that
is considered characteristically normal and non-neoplastic. Our
focus has actually been to look for a true representation through
postmortems. We are just in the process of developing a postmortem
sample-collection process, which is the only true approach to getting
Affiliating with Ardais allowed Duke to have a quality resource
for its own investigators without footing the entire bill, Duke’s
Dr. Proia says. "It’s pretty common to have multiple tissue banks
at an institution—maybe a lung person who collects lung tissue,
and so on," he says. "It’s part of the problem at most academic
places that there are all these little repositories, and the deans
hoped to set up a one-stop shop, so if you had IRB approval to do
research you’d be able to call up and say, ’I need 10 prostates,
20 livers, and are they in stock?’ That was the goal—to have
a comprehensive tissue bank with informed consent, with appropriate
clinical data, with a great diversity of tissues, and the only way
to fund it was to do it in collaboration because it’s horrendously
In the old system, Dr. Proia says, the investigators got tissue
without much data. "Now, because of Duke’s affiliation with Ardais,
I have two research nurses who meet with all patients scheduled
for surgery, and if the surgery is likely to yield bankable tissue,
they ask for their consent, explain that we’re affiliated with Ardais,
that it’s a for-profit company, and there are all these provisions
to ensure their care won’t be compromised."
More than 99 percent of the subjects will consent, Dr. Proia says,
"because medical research has everything to gain and they have nothing
to lose." In 2001, about 900 patients gave their consent for tissue
to be collected and Duke was able to bank approximately half of
it; in 2002, the number of consenting patients rose to over 1,400
and Duke was able to bank about 800.
Even now, however, too many people are
collecting," Dr. Wittliff says. "It’s dangerous because
people don’t store tissue and sera in a standardized format. There’s
no standard way to evaluate tissues for research."
In the 1970s, Dr. Wittliff managed a proficiency testing program
for early tamoxifen trials and an international quality assurance
program for clinical trials on estrogen and progestin receptors.
In the 1980s, he started the biorepositories at the Brown Cancer
The CAP established a formal Survey program on estrogen and progestin
receptors around that time, Dr. Wittliff says, and it continued
until about 2000. "Then it stopped, because most of the proficiency
Survey assays were beginning to be performed by immunohistochemistry,
although even then the biorepository helped with standardization.
Now this biorepository is primarily a frozen tissue bank, which
has turned out to be most valuable since the human genome has been
One of the first tumor markers Dr. Wittliff, along with other
investigators, used the tissue bank to look at was the protein product
of the oncogene HER-2/neu, which indicates an aggressive
breast cancer. "We conducted early studies of HER-2/neu
proteins in our patients’ biopsies, and our studies and others helped
define the ranges of expression," he says. "Then Genentech produced
a new drug—which binds to this protein—to treat breast
cancer, and the drug is called Herceptin."
"We also used the biorepository to define ranges for epidermal
growth factor receptors, and a new drug has just been produced called
Iressa, for breast, lung, and other cancers," he says. Another protein
he studied was a new tumor marker called uPA, urokinase type plasminogen
activator. "Our studies and studies of groups in Europe have indicated
this is a prognostic factor in breast and endometrial cancer, perhaps
in other cancers too, and it’s being integrated into new clinical
trials. This is the current approach to cancer therapy, to synthesize
targeted drugs, and this is where the biorepository is incredibly
Dr. Wittliff is even more enthusiastic about the possibilities
for tissue banks since the commercial development of laser capture
microdissection by Arcturus Applied Genomics, Mountain View, Calif.
"What this instrument does is allow one to dissect a frozen tissue
section and—without any destruction—to capture individual
cancer cells and put them in one tube and the normal cells in another,"
Working with Arcturus, the Brown Cancer Center has been developing
gene expression profiles of pure breast carcinoma cells isolated
from sections of the tissues from the biorepositories, using laser
capture microdissection, and correlating these profiles with significant
clinical data on patients’ nodal status, race, therapeutic outcome,
disease-free survival, and overall survival.
In effect, Dr. Wittliff says, "I’m still mining data and using
samples I stored 20 years ago. What’s extraordinary is that the
protocols set up two decades ago have preserved the activity of
the labile RNA. The proteins are still viable—and we can now
perform gene expression studies from messenger RNA we’re extracting
from biopsies that have remained viable for more than 20 years."
The cost of research tissues, however, is
Depending on the type of tissue and amount of clinical information
needed, the expense of a single tissue sample might reach four figures—out
of range for many academic researchers. Asterand announced in August
2002 that it was revamping its price list for common cancer tissue
samples in response to the different needs of academic and commercial
"We recognize that medical research is affected by budgets and
commercial realities just like most other areas of human endeavor,"
says Asterand business development specialist Victoria Blanc, PhD.
Noting that some experiments have more rigorous requirements than
others, she says, "Rather than having one monolithic price for each
kind of tissue, our new prices will reflect the level of quality
assurance required." Common cancer tissue samples such as lung,
breast, and stomach now range from around $100 to several hundred
dollars if detailed pathology reports are required.
A myriad of complications surrounds informed consent as well.
Margery Moogk, MS, says that researchers who request tissue from
Seattle’s nonprofit Northwest Tissue Bank, which she directs, must
sign a statement agreeing that the tissue will not be commercialized.
"So if a biotech company were trying to develop a diagnostic tool
that recognizes liver cancer, they may need to have normal liver
tissue to compare how that diagnostic tool distinguishes between
normal and abnormal cells, but the product may not have liver cells,"
The Cooperative Human Tissue Network funded by the National Cancer
Institute, which distributes about 80,000 specimens per year in
North America through six participating institutions, has adopted
a similar restriction. "We basically make tissue available to people
who call and ask, and pharmaceutical and clinical diagnostics companies
have access as long as they are doing research and not incorporating
the tissues into product development," says Roger Aamodt, PhD, chief
of the NCI’s Resources Development Branch and president of the International
Society for Biological and Environmental Repositories.
While NCI is in close contact with the big pharmaceutical companies,
it has no formal relationship with them. "We feel we’re serving
pretty much a different constituency," Dr. Aamodt says. But commercial
and academic researchers can use NCI’s specimen resource locator
or "tissue expediter," which functions as a single point of contact
for researchers looking for tissues. In many cases, researchers
don’t have any other means of learning about these tissues because
they’re not commercially available.
A more elaborate effort underway at NCI is the Shared Pathology
Informatics Network, or SPIN, which will use state-of-the-art informatics
techniques to establish an Internet-based virtual database. "This
will allow researchers to query a large number of institutions’
electronic clinical information systems and pull out a listing of
what pathology specimens are there that will meet their research
needs," Dr. Aamodt says. "It will be done in a way that totally
protects patient privacy and confidentiality and will essentially
be infinitely expandable."
SPIN, a five-year project now in its second year, is developing
software to give researchers limited access to de-identified patient
data. NCI says the need for such a system has been fueled by the
growing use of tissues and diagnostic specimens and their related
clinical data in biomedical research.
Since most pathology laboratories store at least 10 years of pathology
reports electronically, there is a wealth of archived tissue and
searchable databases with patient data. SPIN will not create a central
database but will facilitate communications among disparate computer
systems, even among those that use different architectures and search
In the meantime, debate persists over how much regulation the
tissue banking industry needs. "We’re not governed by any overall FDA regulations,"
Asterand’s Charlton points out. "In the U.S., we operate under the rules of
each institution we deal with, whether it’s an academic university or a private
hospital or the federal rules for institutional review boards, and we have our
own as well that meets from time to time. It’s a completely different area from
organ transplantation. This material isn’t going into anybody—just into
a test tube."
Does this mean a cursory informed consent is adequate? The courts
have indicated, Moogk says, that "if a company knows in advance
it will use tissue recovered from a patient’s body and that it will
be a profit-generating thing, you should know that.
"But if an organization is investing a lot of money in research
or new product development, and they don’t have a particular target
in mind, they’re using the tissue without any real knowledge of
or guarantee that it will make a contribution, then it’s not so
clear that they have any kind of obligation to go back to an individual
and say, ’We extracted DNA from your cells that we used to make
the target RNA that is now an important component of our product’—because
it’s not really theirs anymore."
"Law regarding ownership of specimens is very complicated and
confusing at this point," Dr. Aamodt says. The California Supreme
Court, in Moore v. Regents of the University of California,
ruled in 1990 that a patient does not have a continuing ownership
interest in his excised cells and tissue used in research. However,
the court also held that the failure to inform a patient that his
collected tissue would be used for research purposes was a breach
of the duty to obtain informed consent. This is the only published
case on the issue of ownership of excised cells, and there is no
overarching federal rule regarding ownership. In some other states,
diagnostic specimens are considered part of the clinical record.
For tissue that is "unlinked" (rendered anonymous), the CAP opposes
hampering research by well-intended but intrusive regulations. In
November 2001, the CAP’s Ad Hoc Committee on Tissue and Organ Procurement
took the position that consent forms should use simple (general
or unspecified) wording to include donation of excess human tissue
for research, teaching, and quality control testing, and that previously
collected specimens for which no consent form exists should be grandfathered
so they can be available for research. However, because of the evolving
nature of tissue procurement and informed consent laws, the ad hoc
committee is revisiting this position.
Many tissues cannot be "anonymized" because they must be linked
back to clinical information from the patient to allow outcomes
research. In these cases, the CAP cites the federal Office for Human
Research Protection’s requirements for specific types of patient
consent, including education about the operation of the tissue repository,
specific information about the type of research conducted, conditions
under which data or specimens might be released, and procedures
to protect privacy and confidentiality.
"The federal rule that applies, 45CFR46, has a provision that
if there’s no way to identify a subject, you can waive consent with
the approval of the IRB," Dr. Proia says. "We do have a general
consent form with a sort of nebulous provision for materials to
be used for research. That’s where a lot of institutions get into
conflict, because the government says that for federally funded
research you need fully informed consent, but others say the consent
form for surgery allows them to use tissue." So there’s tension,
he says, between the CFR, federally sponsored research requirements,
and the desire of investigators to get tissue when they may not
have the resources to obtain informed consent.
Five years ago, before Duke’s affiliation with Ardais, Dr. Proia
recalls, "some investigators who may have gotten a piece of tumor
from us would call up and say, ’We have interesting results with
this tissue and we need followup.’ And we’d say, ’Didn’t you notice
when you got it there was nothing on it that allowed us to track
the patient down?’ That was part of the stimulus to affiliate,"
he adds, "because the ideal is to have a full repertoire of clinical
data, pathology data, and followup data, and to do that you have
to have fully informed consent from patients undergoing surgery."
For that reason, Dr. Proia says, "we do not collect tissues from
autopsies because of the concern about approaching families after
someone has died." While there have been discussions about piggybacking
onto organ donor programs to request tissue, "due to the whole medicolegal
climate, we haven’t gone there." Duke is considering a rapid autopsy
program, however, in which people would give consent before death.
"You would sign up to be a tissue donor before the research; that
way it would be clean," he says.
More rigorous rules will take effect April 14, Dr. Aamodt says.
"The regulation of tissue banking is the same as rules applying
to any other human subject research, and the Department of Health
and Human Services is about to implement new rules under the Health
Insurance Portability and Accountability Act. These rules will significantly
strengthen privacy and confidentiality controls—probably with
some major negative impact on research," he says.
Most tissue research is considered to pose "minimal risk" to patients,
but the new privacy rules have extensive administrative requirements
that apply whether or not there is a determination of increased
risk. If it is health data and is identifiable, then it requires
additional administrative procedures.
"Because the rules are very complicated, it will take the research
community some time to fully understand what they mean and how they
apply," Dr. Aamodt says. In the meantime, "research institutions
are struggling to get policies to assure confidentiality while allowing
their research to go forward."
The International Society for Biological and Environmental Repositories,
to which many research tissue banks belong, is now organizing standards
for quality control and best practices, perhaps leading to a formal
accreditation process. The group has already created subcommittees
to work on best practices documents in sample collection, processing
and retrieval, sample storage, sample tracking, sample packaging
and shipping, biological safety, training, QA/QC, and human subjects.
"For research tissue banks, there’s no accreditation now, but
it’s probably going to happen," Ardais’ Dr. Buckler predicts. "As
the use of these resources gets closer to the rationale for clinical
development of new therapeutic strategies, it’s going to be a necessary
thing." Ardais is prepared, he adds, because of systems it has already
put in place to document the validity of samples and strict safety
Dr. Wittliff emphasizes that the pathology profession must prepare
as well. "Biorepositories are really a growing resource. But we’ve
got to have a change in the culture of the surgery theatre and in
the pathology suite because the tissues have to be handled differently
than we are accustomed to for routine pathology. Clinical followup
has to be collected, and patient identity must be secure."
Pathologists must be aware of the responsibilities and opportunities
that accompany this new role in tissue banking, Dr. Wittliff says.
"In this post-human genome era, with breakthrough technologies for
proteomics, genomics, and metabolomics, these are the resources
that are necessary. This is where it’s going—and we in pathology
and laboratory medicine must be guardians of the anonymity of our
patients and the integrity of their tissues for research."
1. National Cancer Institute Specimen
Resource Locator: www.cancer.gov/specimens
2. NCITissue Expediter: www.cancerdiagnosis.nci.nih.gov/specimens/finding.html#expediter
3. NCI Cooperative Prostate Cancer Tissue
4. International Society for Biological
and Environmental Repositories: www.isber.org
5. Shared Pathology Informatics Network
Anne Paxton is a writer in Seattle.