ISIS Report 18/03/09

The Golden Rice Scandal Unfolds

Phase II clinical trials on children have been conducted with unapproved experimental GM rice enhanced in pro-Vitamin A that has the potential to cause birth defects and developmental abnormalities Dr. Mae-Wan Ho and Prof. Joe Cummins

This report has been sent to the United States Food and Drug Administration on behalf of ISIS

Clinical trials of unapproved, uncharacterized GM rice on children

More than 30 senior scientists and academics signed an open letter condemning

researchers at Tufts University (Boston) in the United States for carrying out clinical trials of genetically modified (GM) ‘Golden Rice’ on children [1] (Scientists Protest Unethical Clinical Trials of GM Golden Rice). The scientists claimed that the trials were in breach of the Nuremberg Code of Ethics brought in at the end of the Second World War to prevent any repetition of the experiments conducted by Nazi scientists including many on children.

The ‘Golden Rice’ in the trials (GR2) appears to be an experimental collection of transgenic events still in the laboratory, uncharacterised in terms of basic molecular genetics or biological and biochemical properties, not tested pre-clinically on animals, or subjected to any other safety assessment.

Of the three studies listed on the US Clinical Trials website, two involved children. The first, “Retinol Equivalence of Plant Carotenoids in Children” [2] - a phase II trial to compare the vitamin A value of b-carotene in oil capsule, spinach and Golden Rice - recruited 72 children 7 to 9 years of age, and the start and finish dates were September 2004 and November 2005 respectively. The second study, with the same title and also a phase II trial [3], recruited 72 children 6 to 8 years of age and registered start and finish dates July 2008 and January 2009 respectively. No results are available for either of the studies (as of 17 March 2009).

The third study [4], “Bioavailability of Golden Rice Carotenoids in Humans”, listed as “ongoing, but not recruiting participants”, was a phase I trial on 6 adults 40 to 70 years of age. The start date was August 2004, and estimated completion date August 2008. Again, no results are available; though an article in Science (25 April 2008) [5] mentioned “a recent, soon-to-be-published study among healthy volunteers who ate cooked golden rice, led by Robert Russell of Tufts University in Boston.”

All the studies were done in the United States, though it appears that trials were also carried out and/or intended elsewhere.

The Golden Rice Project website [6] (accessed 17 March 2009), stated that “Golden Rice has gone through many tests since it was first obtained” Nine items are listed; but no feeding trial on animals among them. The final two items are:

“8. Tests for beta-carotene bioavailability and bioconversion to retinol….with deuterium-labelled Golden Rice fed to adults in USA and a small group of children in China have been conducted. The former were highly successful and the latter are being evaluated at present.

9. Feeding trials with human adults in China were carried out to measure the effect of fat in the diet, on bioconversion and bioavailability.”

An Indian newspaper reported [7] that a clinical trial was cut short in China in July 2008, when the government found that 24 children 6-8 years of age at a primary school in Henyan, Hunan, were to be used as guinea pigs for a trial with Golden Rice. The trial was sponsored by Tufts University and obtained approval from the US National Institute of Health, though not from the Chinese government, which was alerted by Greenpeace. Greenpeace has also warned the governments of Bangladesh, India, Indonesia, Philippines and Vietnam against the risky trials. But at least three state-owned labs in India are conducting research into Golden Rice: the Indian Agricultural Research Institute in New Delhi, Tamil Nadu Agricultural University, and Hyderabad-based Directorate of Rice Research.

Although Golden Rice was created by Ingo Potrykus at the Institute of Plant Sciences in the Swiss Federal Institute of Technology and Peter Beyer at the University of Freiburg some ten years ago [8], it has remained in the laboratory since. Speaking to Science journal in April 2008 [5], Potrykus bitterly blamed “2 decades of fear-mongering by organizations such as Greenpeace” that has created a regulatory climate so burdensome that only big companies can afford to get any GM products approved. Closer to the truth may be that the product was never ready for commercial approval.

According to a recent report [9], a sample of the Golden Rice grains was sent to Germany in 2001 for a feeding trial with mice. But when the grains were tested for carotenoid content, the scientists were “surprised to find it contained less than one percent of the amount expected.” After the rice was cooked, this was reduced by another 50 percent, so the trial was abandoned.

In 2005, Syngenta made GR2 [10] using the maize version of the enzyme phytoene synthase that was transferred from daffodil. GR2 produced up to 23 times the amount of carotenoids in the original Golden Rice, GR1.

But GR2 was not a transgenic variety based on a single transformation event. On the contrary, it was explicitly stated that [10]: “The reported transgenic rice events [emphasis added] are experimental.” There is no telling whether all the children or adults taking part in any of the trials were given Golden Rice from the same GR2 event. The results of the trials, as yet unreleased, could well be utterly worthless.

Syngenta was donating these GR2 events, via the Humanitarian Project for Golden Rice, for further research and development (to institutes across China, India, Philippines, Indonesia, Bangladesh and Vietnam) “through license under certain conditions”, which include “being governed by the strategic direction of the Golden Rice Humanitarian Board” Requests were to be directed to Adrian Dubock, a previous employee of Syngenta.

Dubock helped Potrykus and Beyer work out a deal in which Syngenta could develop Golden Rice commercially, but farmers in developing countries who make less than US$10 000 a year could get it for free [5]. Dubock retired from Syngenta in 2007, but remains a member of the Golden Rice Humanitarian Board, chaired by Potykus.

Golden Rice, an exercise in how not to do science

Golden Rice, genetically modified to make pro-vitamin A in the endosperm (the grain remaining after polishing), was announced with great fanfare in 2000 as a cure for widespread vitamin A deficiency in developing countries.

The project had already cost US$100 million, funded by the Rockefeller Foundation, the Swiss Federal Institute of Technology, the European Community Biotech Programme and the Swiss Federal Office for Education and Science, and could cost as much again to develop. It was tied up in at least 70 patent claims on genes, DNA sequences and constructs, a problem only partly solved in the “ground-breaking deal” worked out by Dubock (see above)..

Condemnation was swift and widespread, not least because it was absurd to offer Golden Rice as the cure for vitamin A deficiency when there are plenty of alternative, infinitely cheaper sources of vitamin A or pro-Vitamin A, such as green vegetables and unpolished coloured rice (especially black and purple varieties [11], which would be rich in other essential vitamins and minerals, and hence much more nutritious. The UN Food and Agricultural Organization (FAO) started a project in 1985 to deal with vitamin A deficiency using a combination of food fortification, food supplements and general improvements in diets by encouraging people to grow and eat a variety of green leafy vegetables. One main discovery from the project was that the absorption of pro-vitamin A depends on the overall nutritional status, which in turn depends on the diversity of the food consumed [12].

The main cause of hunger and malnutrition in the Third World is the industrial monocultures of the Green Revolution, which obliterated agricultural biodiversity and soil fertility, resulting in ever-worsening mineral and micronutrient deficiencies in our food. Golden Rice, like other GM crops, is industrial monoculture only worse, and will exacerbate this trend, as well as the destruction of agricultural land, and the impoverishment of family farmers that also accompanied the Green Revolution [13] (see Beware the New "Doubly Green Revolution", SiS 37).

GR1 was made with the standard ‘first generation’ genetic modification techniques, using GM constructs that cause uncontrollable mutations and other collateral damage to the host plant genome, with many unintended, uncharacterized effects [14]. In addition, the viral and bacterial sequences, including antibiotic resistance marker genes, in the construct and in the vectors created for gene transfer enhance horizontal gene transfer and recombination, the main route to creating new pathogens and spreading antibiotic resistance.

GR2 represents an improvement in so far as antibiotic resistance markers were no longer used, but still includes a medley combination of sequences from plant pathogens Agrobacterium (used in a binary vector system) and Erwinia uredovor, and from E. coli, inhabitant of the human gut, which also contains pathogenic strains. We have highlighted the special hazards of the Agrobacterium vector system since 2003 [15] (Agrobacterium & Morgellons Disease, A GM Connection?, SiS 38) (see below).

The main reason for Golden Rice was revealed in the unusually long news feature article [16] accompanying the scientific publication [8] which stated: “One can only hope that this application of plant genetic engineering to ameliorate human misery without regard to short-term profit will restore this technology to political acceptability.”

A detailed audit on the project [14] (The 'Golden Rice', An Exercise in How Not to Do Science, ISIS Report) uncovered “fundamental deficiencies” from the scientific and social rationale to the science and technology involved. It was being promoted “to salvage a morally as well as financially bankrupt agricultural biotech industry.”  The situation has changed little since.

The phase II clinical trials of uncharacterized, unapproved, experimental GR2 events on children, some of whom may indeed be suffering from vitamin A deficiency, is morally inexcusable. GR2 has not been assessed for safety, and there are reasons to suspect it is unsafe.

GMO safety in question

The biotech industry has consistently found genetically modified food and feed ‘as safe as their conventional counterparts’, and regulators in the United States and European Union have accepted this assertion overwhelmingly based on studies carried out and interpreted by the industry [17] (GM Food Nightmare Unfolding in the Regulatory Sham, ISIS scientific publication).

There is now a string of evidence that exposure of many species of animals to a variety of genetically modified crops, and food and feed derived from them, can cause illnesses and death, raising the distinct possibility that genetic modification is inherently dangerous [18] (GM is Dangerous and Futile, SiS 40). This is reinforced in results obtained in the most recent studies.

The Austrian government commissioned long term studies showing that mice fed GM maize hybrid (NK603xMON810) with combined glyphosate tolerance and biopesticide Cry1Ab produced fewer and smaller litters with many genes affected compared to controls [19] (GM Maize Reduces Fertility & Deregulates Genes in Mice, SiS 41). At the same time, the Italian National Institute of Research published a study showing that GM maize MON810 fed to mice produced disturbances in the immune system of the young and the old [20] (GM Maize Disturbs Immune System of Young and Old Mice, SiS 41). In India, the first independent assessment of the feeding study submitted by Monsanto and its subsidiary Mahyco to the Indian regulatory authorities showed that Bt Brinjal (aubergine) caused many changes in several species of animals including diarrhoea, increased water consumption and decrease in liver weight in rats [21] (Bt Brinjal Unfit for Human Consumption, SiS 41).

There are several reasons why genetic modification is inherently hazardous, as spelt out more than ten years ago [22] (Genetic Engineering: Dream or Nightmare?) and unfortunately, still not taken on board by the regulatory authorities, let alone systematically investigated. The dangers may come from the transgenic protein itself that may be toxic or immunogenic [23] (Transgenic Pea that Made Mice Ill, SiS 29), the toxicity of herbicides such as glyphosate to which more than 70 percent of GM crops now grown globally are made tolerant [24] (Death by Multiple Poisoning, Glyphosate and Roundup, SiS 42) or it could be totally unexpected, unintended effects resulting from the mutagenic insertion of foreign DNA into the genome, and worse, the instability of transgenic lines, which makes proper safety assessment well nigh impossible [25] (Transgenic Lines Unstable hence Illegal and Ineligible for Protection, SiS 38).

One major hazard inherent to GM organisms (GMOs) is enhanced horizontal gene transfer and recombination [26] (Horizontal Gene Transfer from GMOs Does Happen, SiS 39). This is considerably worse with transgenic plants like Golden Rice (both GR1 and GR2) that have been created using the Agrobacterium binary vector system, basically because the Agrobacterium bacteria as well as the binary vector tend to persist in the transgenic plants, providing a ready vehicle for further horizontal gene transfer to all species that interact with the transgenic plant material, including human cells. Agrobacterium is known to invade human cells. Horizontal transfer of transgenic DNA into human cells has the potential to cause harmful mutations including cancer. In general, horizontal transfer of transgenic DNA facilitates the creation of new pathogens. The identification of Agrobacterium sequences in patients with Morgellons’ Disease raises questions as to whether the widespread use of Agrobacterium vectors in genetic modification has indeed resulted in creating a new pathogen for humans [15].

Golden Rice particularly dangerous

In addition, the unbalanced enhancement of single nutrients in GM crops may do more harm than good [27] (GM Crops and Microbes for Health or Public Health Hazards?  SiS 32). As David Schubert at the Salk Institute for Biological Sciences La Jolla, California, in the United States points out [28], plants possess the ability to synthesize between 90 000 and 200 000 nonessential small molecules, with up to 500 in one species. The enormous repertoire is due in part to enzymes with very low substrate specificity, which are unpredictably altered by mutations and pleiotropic effects associated with GM technology. Furthermore, overdose of many single nutrients are known to be toxic, vitamin A being a case in point. Schubert highlights the toxic effects of retinoic acid and other metabolites of b-carotene, only a few of them can be identified and measured in the current state of technology.

Golden Rice is enhanced in b-carotene, which on ingestion, is cleaved in half to generate retinal for use in the visual cycle. Retinal is also reduced to retinol, or oxidized to retinoic acid (RA), which interacts with highly specific nuclear receptors. Essentially all of the biological activity of retinoids, apart from vision, involves RA. While high concentrations of retinol are toxic, RA is biologically active at concentrations several orders of magnitude lower than retinol. Hence, Schubert states [28]:  “excess RA or RA derivatives are exceedingly dangerous, particularly to infants and during pregnancy.” RA is required for the development of the nervous system, both by directly controlling nerve differentiation and by generating concentration gradients that direct cell migration, embryonic segmentation, and development. Therefore, RA and synthetic derivative of RA are teratogenic (able to cause birth defects). They can accumulate in fat and plasma, becoming a risk factor for pregnancy for up to 2 years following ingestion, and multiple low doses of retinoids have greater toxicity than a single high dose.

Because of the type of biological functions controlled by low levels of RA, any perturbation of its signalling pathways by plant-derived RA receptor agonists or antagonists will have clinical consequences. “Could the GM modifications used to enhance b-carotene synthesis create such compounds?” (This question remains unanswered to this day.) Six hundred naturally occurring compounds exist in the carotene family, and at least 60 can be precursors to retinoids. “Therefore, plants have the potential to make many potentially harmful retinoid-like compounds when there are increased levels of synthetic intermediates to b-carotene as in golden rice.”

While all retinoids and derivatives are likely to be teratogenic, good assays and information regarding the behaviour and teralogic activity are available for only three: retinol, RA, and retinal. Therefore, at the very least, “extensive safety testing should be required before the introduction of golden rice as a food.”

References

1. Scientists Protest Unethical Clinical Trials of GM Golden Rice, Open Letter, 12 February 2009, for complete list of signatories see http://www.gmfreecymru.org/open_letters/Open_letter12Feb2009.html)
2. Project NCT 00082420. Retinol Equivalence of Plant Carotenoids in Children. http://clinicaltrials.gov/show/NCT00082420
3. Project NCT 00680212. Vitamin A Equivalence of Plant Carotenoids in Children. http://clinicaltrials.gov/ct2/show/record/NCT00680212?term=golden +rice&rank=3
4. Project NCT 00680355.(10) Bioavailability of Golden Rice Carotenoids in Humans. http://clinicaltrials.gov/ct2/show/record/NCT00680355?term=golden +rice&rank=1
5. Enserink M. Tough lessons from Golden Rice. Science 2008, 320, 468-71.
6. Research on Biosafety, http://www.goldenrice.org/Content2-How/how3_biosafety.html
7.  “A new genetically modified rice strain is breeding controvery”, Noemie Bisserbe 26 August 2008, Businessworld, http://www.businessworld.in/index.php/Economy-and-Banking/Golden-Scare.html
8. Ye X, Al-Babili S, Kloti A., Zhang J, Lucca P, Beyer P. and Potrykus I. Engineering the provitamin A (b-carotene) biosynthetic pathway into (carotenoid-free) rice endosperm. Science 2000, 287, 303-5.
9. Then C. A critical look at golden Rice after nearly 10 years of development. C Commissioned by Foodwatch in Germany, January 2009.
10. Paine JA, Shipton CA, Chaggar S, Howells RM, Kennedy MJ, Vernon G, Wright SY, hinchliffe E, Adams JL, Silverstone AL and Drake R. Improving the nutritioinal value of Golden Rice through increased pro-vitamin A content. Nature biotechnology 2005, 21, 482-7.
11. Frei M and Becker K. Fatty acids and all-trans-b-carotene are correlated in different colored rice landraces. J Sci Food Agri  2005, 85, 2380-4.
12. Koechlin F. The ‘golden rice’ – a big illusion? Third World Resurgence 2000, #114/115, 33-35.
13. Ho MW. Beware the new “doubly Green Revolution”. Science in Society 37, 26-29, 2008.
14. Ho MW. The ‘Golden Rice’ – An Exercise in How Not to Do Science. TWN Biotechnology and Biosafety Series No. 6, Third World Network, Penang, 2002. http://www.i-sis.org.uk/onlinestore/books.php#276
15. Ho MW and Cummins J. Agrobacterium & Morgellons Disease, a GM connection? Science in Society 38, 33-35, 2008.
16. Guerinot ML. The Green Revolution strikes gold. Science 2000, 287, 241-3.
17. Ho MW, Cummins J and Saunders PT. GM food nightmare unfolding in the regulatory sham. Microbial Ecology in Health and Disease 2007, Disease 2007, 19, 66-77. http://www.i-sis.org.uk/onlinestore/papers2.php#section5
18. Ho MW. GM is dangerous and futile, we need organic sustainable food and energy systems now. Science in Society 40, 4-8, 2008.
19. Ho MW. GM maize reduces fertility & deregulates genes in mice. Science in Society 41, 40-41, 2009.
20. Ho MW. GM maize disturbs immune system of young and old mice. Science in Society 41, 42, 2009.
21. Burcher S. Bt brinjal unfit for human consumption. Science in Society 41, 50-51, 2009.
22. Ho MW. Genetic Engineering Dream of Nightmare? The Brave New World of Bad Science and Big Business, Third World Network, Gateway Books, MacMillan, Continuum, Penang, Malaysia, Bath, UK, Dublin, Ireland, New York, USA, 1998, 1999, 2007 (reprint with extended Introduction). http://www.i-sis.org.uk/onlinestore/books.php#253
23. Ho MW. Transgenic pea that made mice ill. Science in Society 29, 28-29, 2006.
24. Ho MW and Cherry B. Death by multiple poisoning, glyphosate and Roundup. Science in Society 42 (to appear).
25. Ho MW. Transgenic lines unstable hence illegal and ineligible for protection. Science in Society 39, 28-29, 2008.
26. Ho MW and Cummins J. Horizontal gene transfer from GMOs does happen. Science in Society 39, 22-24, 2008.
27. Cummins J and Ho MW. GM crops and microbes for health or public health hazards? Science in Society 32, 30-33.
28. Schubert DR. The problem with nutritionally enhanced plants. J Medicinal Food 2008, 11, 601-5.

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