COLUMN ONE : The Mouse Wars Turn Furious : It’s now so easy to genetically engineer lab animals that everyone, it seems, is out to patent the perfect specimen. The explosion in knowledge has also spawned charges of greed, exploitation.

At first glance, the rat bears a striking resemblance to its common cousins that scramble among the dust balls of abandoned buildings. But on closer examination, there is something odd about this rodent. Oddly human, to be exact.

Its limbs are swollen with arthritis. It has the dry, flaky skin of psoriasis and the chronic upset stomach that comes with colitis.

Such astonishing afflictions make the HLA-B27 rat--created at the University of Texas, raised in Upstate New York and marketed by a San Francisco-area biotechnology firm, GenPharm International Inc.--a valuable commodity. A patent is pending for these rodents, which cost $285 each and have an electronic bar code implanted under their skin to allow easy identification.

Around the world, so many animals like this un-ratlike rat have been stricken with human diseases that they would fill a dozen hospital wards if they were human. Among them: Rabbits in Massachusetts are sick with the AIDS virus, rodents in Berkeley suffer sickle cell anemia, and mice in North Carolina have hardened arteries and cystic fibrosis.

Still other creatures borrow human physiology. In Ohio, pigs are born with human-like blood and perhaps even organs that might pass as human, and dairy cows in the Netherlands produce milk that contains the same therapeutic proteins found in a woman’s breast milk.

Freaks of nature? Far from it. These are “designer” animals--intentionally created by humans to resemble humans.

Scientists now almost routinely knock out animal genes in an embryo and plunk in human ones, inducing mutations that mimic human traits or maladies. They can, in theory, fashion almost any type of rodent they want.

In effect, the scientists are creating miniature, furry patients to examine some of the world’s deadliest and most baffling maladies, and they are experimenting with turning livestock and rodents into pharmaceutical factories. The new creatures are considered important because they provide living laboratories in which scientists can study diseases that ethically cannot be inflicted on human test subjects.

“This is just the beginning,” said Kenneth Paigen, director of the nonprofit Jackson Laboratory in Bar Harbor, Me., the world’s largest producer of mutated mice. “In terms of our understanding of ourselves and changes in the ways we live and what we will die from, what’s happening now is probably the most important revolution that society has seen.”

But by blurring the lines between people and animals, this latest explosion in genetic engineering is not only transforming medical and developmental biology, it is raising disturbing legal, economic and moral quandaries for scientists.

Everyone, it seems, is clamoring to invent, patent and market a million-dollar lab mouse, and at times the business has degenerated into furious mouse wars.

Accusations of greed and exploitation have spread, and many scientists fear that growing commercialization of their valuable laboratory tool is escalating costs and thereby stifling medical research that could benefit the public.

Also simmering just beneath the surface are vexing questions about the moral propriety of creating, patenting and selling new forms of life. Outraged biotech opponents rebuke it as dangerous tinkering with nature.

“A special case of dispute and controversy surrounds the manipulation of life,” Caltech science historian Daniel Kevles said. “When people talk about manipulating animals and changing their genetic essence, that is different than just breeding animals.”

Mutant Strains

Discovered in 1980 and refined in 1988 to target specific genes, this emerging “transgenic” technology has already spawned a massive, worldwide academic and commercial enterprise, with investments perhaps topping $1 billion.

Hundreds of laboratories are mixing the DNA of humans and animals, and probably at least 5,000 different strains of mutant creatures have been produced in the last few years. So far, most of the DNA swapping has focused on mice because, unlike other mammals, much of their genetic code has already been unraveled.

Four genetically altered mice and one rabbit have been patented, giving their creators exclusive marketing rights, and an estimated 140 other animals have patent applications pending with the U.S. government.

The transgenic animal boom “will be seen historically as one of the major developments in medicine and certainly in genetics,” said Maxwell Cowan, vice president and chief scientific officer of the Howard Hughes Medical Institute, which funds several researchers who helped pioneer the technology as well as dozens who use it.

“We can now produce animals with essentially any genetic disorder,” he said. “We have in hand a potentially enormous tool.”

For the first time, mammals can be produced with a specific human malady, such as HIV-infected rabbits at the National Institutes of Health that bear the human receptor gene that spreads the AIDS virus, mice at Harvard Medical School with enlarged prostates, or GenPharm’s HLA-B27 rat, which mimics the multiple symptoms of an inflammatory disease called spondyloarthropathies.

Researchers can use these animals to explore how puzzling diseases manifest themselves and test potential cures, drugs and other treatments.

“It opens up a whole new phase of biology--the ability to make changes at will in the specific gene of an animal,” said Dr. Oliver Smithies, a University of North Carolina pathologist who was one of the pioneers of genetic targeting techniques and helped create mice with cystic fibrosis and hardened arteries.

“There will be very significant uses of these animals,” he said. “It could be as soon as a year, or as long as five years. But I think it won’t be longer than that before we’ll find something helpful in human disease.”

Major Impact

Before the mid-1980s, medical researchers who wanted to study an organism afflicted with a human disease relied mostly on cloned bacteria in a test tube. But a single-celled E. coli reacts much differently than a mouse, which has much of the same physiology as a human.

“In our case, you can’t look at heart disease in a test tube, so these transgenic animals have a very major impact,” said Edward Rubin, a Lawrence Berkeley Laboratory geneticist who created mice with so much “good” cholesterol that they are nearly immune to heart disease.

“They contribute to our understanding of how things work in a setting fairly close to the way it works in people,” he said. “It’s really been a gold mine for researchers.”

The technique of custom-making a rodent, considered the stuff of science fiction a decade ago, has become almost low-tech. It has become so routine at many laboratories that GenPharm President David Winter calls it “dial-a-mouse.”

“I predict in 10 years that making these animals will be a high school or college experiment,” said Winter, whose company has 10,000 gene-altered mice on hand at any time. “Students getting a master’s will have to knock out genes and put them in.”

First, a scientist isolates and clones a human gene that causes a disease such as cystic fibrosis and inserts it into cells taken from a mouse’s embryo. The human genes target their mouse counterparts, overpowering them.

A technician methodically locates these mutated cells and, using a needle with a hair-thin point and a sophisticated microscope, literally pushes them into another mouse embryo. Three weeks later, a mouse is born with cystic fibrosis or whatever disease has been targeted. Cross-breeding the offspring produces a long lineage of mice with the defective gene.

In one afternoon, a trained biological technician can inject the mutated cells into a couple hundred mouse embryos.

In the last three years, the scene has turned into one of mass production. Laboratories around the world are cranking out tailor-made mice for research, and commercial biotechnology firms such as GenPharm are buying up rights to the creatures and marketing them. Last year, about 700 scientific articles were published with new revelations about transgenic animals.

“Basically the only thing that stops this explosion is funding limitations, because the strains (of mice) are expensive to produce and maintain,” said Eric Fischer, director of the National Academy of Sciences’ Board on Biology.

Marketing gimmicks, complete with catchy names, have emerged. Scientists can call (800) LAB-RATS to take their pick of regular rodents or seven strains of transgenic ones. One firm is named Pig Improvement Co. A prestigious London institute dubbed its rodent “ImmortoMouse.”

Electronic Catalogue

An electronic catalogue is being created at Oak Ridge National Laboratory to help scientists locate the rodent they need. And Jackson Laboratory is gearing up a national repository where altered mouse embryos will be frozen and preserved, just in case ones that seem useless or obsolete today prove valuable tomorrow.

Animal rights activists and genetic engineering opponents in the United States have been unusually quiet during this biotech boom.

But opponents say they are rallying their forces to launch a nationwide political campaign this month to seek controls on the engineering and patenting of animals.

“This whole thing raises profound environmental and economic and social issues--more so than any other technology in history,” said Jeremy Rifkin of the Foundation on Economic Trends, who is perhaps the nation’s most vocal critic of genetic engineering.

“It shows the extremes to which this capitalist system has gone,” Rifkin said. “We are embarking on a very potentially troublesome journey, where we begin to reduce all other animals on this planet to genetically engineered products. . . . We will increasingly think of ourselves as just gene codes and blueprints and programs that can be tinkered with.”

Rifkin said he also fears that new deadly maladies might be unleashed in the animal community. Scientists, however, call such concerns irrational because new animal strains pose no known risk and people have always altered nature.

“Nature is not immutable,” said Jonathan MacQuitty, chief executive officer of GenPharm International, which makes transgenic dairy cows and mice that produce human antibodies. “It changes all the time, constantly, and we have a huge impact on it. People have to realize we are already enmeshed in nature. We shouldn’t pretend we don’t exist.”

Geneticists acknowledge that the animals have not yet borne major medical breakthroughs. In an embarrassing public failure, scientists who initially reported that they had created mice with Alzheimer’s disease had to retract their findings, and other researchers remain slightly off the mark in mimicking various diseases.

A Question of Profit

Still, physicians say these gene-altered rodents are the best hope they have.

“These are reasonably good working models of disease in man. They don’t always exactly emulate it, but we have little choice,” said Dr. Harold Strauss, a Duke University professor of medicine who serves on the American Heart Assn.’s board of directors. “It gives us tremendous hope for breakthroughs in the future.”

Most of the concerns have been economic rather than scientific, with some researchers accusing biotechnology companies of charging exorbitant prices.

Most scientists abhor secrecy and proprietary rights in science, and they demand an open exchange, including free or cheap access to laboratory tools such as the mice. But private companies and some researchers and universities say they cannot justify spending millions creating and breeding the animals for other scientists without protecting their investment.

It is a fine line, researchers say, between fostering technology and exploiting it, and they worry that the transgenic animal business is teetering on the edge.

“We have just passed through a period of exaggerated greed and exaggerated expectations,” said Cowan of the Howard Hughes Medical Institute, “but I think this will all shake out.”

Patent Issue Divisive

No issue has been as divisive as patents. Should government grant the same creation rights for animals as it does inanimate objects like microwaves and light bulbs?

The legal aspect of the patent question has already been addressed. In 1980, the U.S. Supreme Court ruled that man-made living organisms should be issued patents like any other invention. Patents grant exclusive rights to create, use and market a creation for 17 years.

Harvard obtained the nation’s first animal patent in 1988 for the OncoMouse, a cancer-prone rodent. Then the U.S. Patent and Trademark Office, facing an escalating backlog of similar animals and intense public opposition, took almost five years to issue the next ones. In December, patents were awarded to Harvard, GenPharm and Ohio University for gene-altered mice.

Then, three months ago, the National Institutes of Health quietly obtained a patent for rabbits that are genetically engineered to contract the human immunodeficiency virus, which causes AIDS.

A notorious and widely despised Harvard-Du Pont agreement is often cited as evidence of how animal licensing has gone awry. Harvard sold the rights for its patented OncoMouse to Du Pont, which tacked on heavy-handed restrictions that limit its use and require royalties. Scientists have largely boycotted the mouse.

Although Du Pont has not budged, GenPharm decided to change its contracts last fall after facing the wrath of the scientific community for similar policies.

The company now has a one-time “use fee” of $80 to $285 per mouse with no royalties or restrictions. Nonprofit scientists who want to breed GenPharm’s mice must pay $1,000 per year, and a private company must pay $10,000 annually.

Some of the nation’s most prestigious universities--including Massachusetts Institute of Technology, Harvard, Columbia and Rockefeller--have signed contracts with commercial companies to sell their gene-altered animals.

“We’re not going to allow rapacious pricing or restrictions. But the fact that it is a profit-making entity does not seem to us to be against the American way,” said Lita Nelsen, licensing director at MIT, which created a mouse with an altered immune system that is sold exclusively by GenPharm.

Still, many scientists complain that the prices charged by the companies are a rip-off. Some refuse to go commercial, spurning the contracts as if they were a license with the devil.

Rubin of Lawrence Berkeley Laboratory said GenPharm offered to pay his laboratory in exchange for rights to market his high-cholesterol mouse, but he declined because he was afraid that the company would gouge his colleagues. Instead, he breeds them himself and ships them to his fellow scientists.

“I think GenPharm does interesting things,” Rubin said, “but they are a company that wants to sell mice that other people have created and make a profit being the middleman.”

Expensive Effort

Mouse-marketing businesses say the real greed is on the part of some universities, which have set up their own licensing offices. One major university recently contacted GenPharm, offering to sell rights to a mouse for $250,000 plus 15% of any royalties and $20,000 in annual fees. The company refused, saying the mouse was not worth it.

The companies say their prices are justified because mass production is financially risky, expensive and time-consuming. Developing one colony costs an estimated $90,000 and takes two years, according to GenPharm.

“It’s very expensive to keep animals disease-free and keep them healthy and ship them,” said Glenn Monastersky, transgenics director of Charles River Laboratories, which sells seven strains of transgenic mice. “If these animals are going to be used worldwide, these researchers need people like us to distribute them.”

Some of the anxiety might soon be relieved because Jackson Laboratory in Bar Harbor, Me., plans to open a nonprofit national repository to distribute the mice at cost. In all likelihood, however, the ones with patents and other exclusive licenses will not be available.

Mutant mice so far have been a money-loser for private companies. GenPharm, which has $27 million in equity, has invested about $4 million in genetically altered mice in the last two years. Orders have been placed by 140 customers, with a typical order of $400 to $600 for five mice. At that rate, it will take years for GenPharm to pay off its expenses.

Of course, these companies wouldn’t have leaped into the new mouse business if they didn’t expect to turn a profit eventually. They hope pharmaceutical firms will begin buying thousands of diseased rodents to test new drugs, pesticides and other products.

The biggest profits, however, will probably come in a new transgenic industry called “pharming”--genetic engineering of livestock to create pharmaceutical products.

Biotech companies are experimenting with altered mice, pigs, goats and cattle to produce plentiful antibodies, organs or other health care products for people. Still others are working on altering genes to create “super” livestock--leaner cattle, fish that grow bigger or faster, or even sheep that in essence “shear” themselves after being injected with a growth protein that causes their wool to fall off in a single sheet.

New Jersey-based DNX Corp. has created pigs with human hemoglobin and hopes to begin testing the product in humans as a blood substitute next year.

DNX also is trying to develop pigs with hearts, kidneys, lungs and livers that excrete human proteins to disguise the organs so the human body doesn’t automatically reject them. The company hopes to create a large bank of transplantable organs to sell; an estimated 150,000 people die each year because of a worldwide organ shortage.

The lingering questions are not so much in the science, but whether these products can be made economically, said DNX President Paul Schmitt. The organ project, for example, could cost $50 million to $100 million.

“We’ve all got a long way to go,” he said, “but now it’s the nuts and bolts of grinding out product development. Just think about what we’re dealing with. People are sitting around dying because there aren’t enough organs. We are addressing desperate medical needs.”

Biotech Explosion

Transgenic technology could theoretically evolve to the point of engineering totally new life forms.

Today’s scientists know how to alter only a minuscule fraction of an animal’s DNA--at most 0.03%, even with a new technique announced in March that enables researchers to insert 20 times more DNA into a mouse.

“Once you have the ability to add and subtract genes from a mouse and reconstruct a human genetic trait, the possibilities are enormous,” said Paigen, director of the Jackson Laboratory.

So many tailor-made rodents have been produced that scientists can’t keep track of which ones exist and who owns them, and strains are constantly being “discontinued”--dying off with no one bothering to keep the formula.

“Things are happening so quickly with transgenics that we need a very efficient and quick electronic means of tracking them and disseminating the information,” said Richard Woychik, a senior research scientist at the Oak Ridge National Laboratory in Tennessee, which is establishing a computer database for mutated mice.

Scientists look forward to flipping through a computerized catalogue, perhaps even custom-ordering the mouse they want.

“At some time down the line, you’ll be able to go to a catalogue and say, ‘OK, I want a mouse that makes a lot of this particular gene,’ ” Rubin said. “I think that is the future. These transgenic mice are going to be tools of science for a very long time.”

Creating a New Mouse

Biologists are creating thousands of strains of genetically mutated animals by injecting them with human DNA. This technique, called transgenics, allows them to produce mice or other animals afflicted with various human diseases, such as cystic fibrosis or AIDS, to experiment with treatments and cures. Scientists can even target a specific gene to replace. Here’s how it works:

1. Remove embryo from white mouse 3 1/2 days after fertilization. From embryo, collect stem cells (individual cells each capable of creating an entire animal.) Put cells in culture dish.

2. Select human gene that causes disease and isolate its mouse cell equivalent. Hit mouse’s stem cells with electrical pulse that perforates cell’s membrane. The human DNA mutates mouse DNA, causing cell defect similar to human disease.

3. Identify cells that incorporated human DNA and clone them.

4. Remove second embryo from black mouse.

5. Using microscope and needle, push mutated cells into embryo of black mouse.

5. Implant embryo in “surrogate” mouse.

6. In about three weeks, the new litter will be speckled black and white if human DNA was they successfully incorporated.

Source: GenPharm International Inc.