Differences in the carcinogenic evaluation of glyphosate between the International Agency for Research on Cancer (IARC) and the European Food Safety Authority (EFSA)

The International Agency for Research on Cancer (IARC) Monographs Programme identifies chemicals, drugs, mixtures, occupational exposures, lifestyles and personal habits, and physical and biological agents that cause cancer in humans and has evaluated about 1000 agents since 1971. Monographs are written by ad hoc Working Groups (WGs) of international scientific experts over a period of about 12 months ending in an eight-day meeting. The WG evaluates all of the publicly available scientific information on each substance and, through a transparent and rigorous process,¹ decides on the degree to which the scientific evidence supports that substance's potential to cause or not cause cancer in humans.

For Monograph 112,² 17 expert scientists evaluated the carcinogenic hazard for four insecticides and the herbicide glyphosate.³ The WG concluded that the data for glyphosate meet the criteria for classification as a probable human carcinogen.

The European Food Safety Authority (EFSA) is the primary agency of the European Union for risk assessments regarding food safety. In October 2015, EFSA reported⁴ on their evaluation of the Renewal Assessment Report⁵ (RAR) for glyphosate that was prepared by the Rapporteur Member State, the German Federal Institute for Risk Assessment (BfR). EFSA concluded that ‘glyphosate is unlikely to pose a carcinogenic hazard to humans and the evidence does not support classification with regard to its carcinogenic potential’. Addendum 1 (the BfR Addendum) of the RAR⁵ discusses the scientific rationale for differing from the IARC WG conclusion.

Serious flaws in the scientific evaluation in the RAR incorrectly characterise the potential for a carcinogenic hazard from exposure to glyphosate. Since the RAR is the basis for the European Food Safety Agency (EFSA) conclusion,⁴ it is critical that these shortcomings are corrected.

The human evidence

EFSA concluded ‘that there is very limited evidence for an association between glyphosate-based formulations and non-Hodgkin lymphoma (NHL), overall inconclusive for a causal or clear associative relationship between glyphosate and cancer in human studies’. The BfR Addendum (p. ii) to the EFSA report explains that ‘no consistent positive association was observed’ and ‘the most powerful study showed no effect’. The IARC WG concluded there is limited evidence of carcinogenicity in humans which means “A positive association has been observed between exposure to the agent and cancer for which a causal interpretation is considered by the Working Group to be credible, but chance, bias or confounding could not be ruled out with reasonable confidence.”¹

The finding of limited evidence by the IARC WG was for NHL, based on high-quality case–control studies, which are particularly valuable for determining the carcinogenicity of an agent because their design facilitates exposure assessment and reduces the potential for certain biases. The Agricultural Health Study⁶ (AHS) was the only cohort study available providing information on the carcinogenicity of glyphosate. The study had a null finding for NHL (RR 1.1, 0.7–1.9) with no apparent exposure–response relationship in the results. Despite potential advantages of cohort versus case–control studies, the AHS had only 92 NHL cases in the unadjusted analysis as compared to 650 cases in a pooled case–control analysis from the USA.⁷ In addition, the median follow-up time in the AHS was 6.7 years, which is unlikely to be long enough to account for cancer latency.⁸

The RAR classified all of the case–control studies as ‘not reliable,’ because, for example, information on glyphosate exposure, smoking status and/or previous diseases had not been assessed. In most cases, this is contrary to what is actually described in the publications. Well-designed case–control studies are recognised as strong evidence and routinely relied on for hazard evaluations.^{9 10} The IARC WG carefully and thoroughly evaluated all available epidemiology data, considering the strengths and weaknesses of each study. This is key to determining that the positive associations seen in the case–control studies are a reliable indication of an association and not simply due to chance or methodological flaws. To provide a reasonable interpretation of the findings, an evaluation needs to properly weight studies according to quality rather than simply count the number of positives and negatives. The two meta-analyses cited in the IARC Monograph¹¹ are excellent examples of objective evaluations and show a consistent positive association between glyphosate and NHL.

The final conclusion⁵ (Addendum 1, p.21) that “there was no unequivocal evidence for a clear and strong association of NHL with glyphosate” is misleading. IARC, like many other groups, uses three levels of evidence for human cancer data.¹ Sufficient evidence means ‘that a causal relationship has been established’ between glyphosate and NHL. BfR's conclusion is equivalent to deciding that there is not sufficient evidence. Legitimate public health concerns arise when ‘causality is credible’, that is, when there is limited evidence of carcinogenicity.

Evidence from animal carcinogenicity studies

EFSA concluded ‘No evidence of carcinogenicity was confirmed by the majority of the experts (with the exception of one minority view) in either rats or mice due to a lack of statistical significance in pairwise comparison tests, lack of consistency in multiple animal studies and slightly increased incidences only at dose levels at or above the limit dose/maximum tolerated dose (MTD), lack of preneoplastic lesions and/or being within historical control range’. The IARC WG review found a significant positive trend for renal tumours in male CD-1 mice,¹² a rare tumour, although no comparisons of any individual exposure group to the control group were statistically significant. The WG also identified a significant positive trend for hemangiosarcoma in male CD-1 mice,¹³ again with no individual exposure group significantly different from controls. Finally, the WG also saw a significant increase in the incidence of pancreatic islet cell adenomas in two studies in male Sprague-Dawley rats.^14–16 In one of these rat studies, thyroid gland adenomas in females and liver adenomas in males were also increased. By the IARC review criteria,¹ this constitutes sufficient evidence in animals.

The IARC WG reached this conclusion using data that were publicly available in sufficient detail for independent scientific evaluation (a requirement of the IARC Preamble¹). On the basis of the BfR Addendum, it seems there were three additional mouse studies and two additional rat studies that were unpublished and available to EFSA. Two of the additional studies were reported to have a significant trend for renal tumours, one in CD-1 mice (Sugimoto. 18-Month Oral Oncogenicity Study in Mice. Unpublished, designated ASB2012–11493 in RAR. 1997), and one in Swiss-Webster mice (Unknown. A chronic feeding study of glyphosate (roundup technical) in mice. Unpublished, designated ABS2012–11491 in RAR. 2001). One of these studies (Sugimoto. Unpublished, 1997) also reported a significant trend for hemangiosarcoma. The RAR also reported two studies in CD-1 mice showing significant trends for malignant lymphoma (Sugimoto. Unpublished, 1997; Unknown. Glyphosate Technical: Dietary Carcinogencity Study in the Mouse. Unpublished, designated ABS2012–11492 in RAR. 2009).

The RAR dismissed the observed trends in tumour incidence because there are no individual treatment groups that are significantly different from controls and because the maximum observed response is reportedly within the range of the historical control data (Table 5.3–1, p.90). Care must be taken in using historical control data to evaluate animal carcinogenicity data. In virtually all guidelines,^{1 17 18} scientific reports¹⁹ and publications^20–23 on this issue, the recommended first choice is the use of concurrent controls and trend tests, even in the EC regulations cited in the RAR¹⁸ (see p.375). Trend tests are more powerful than pairwise comparisons, particularly for rare tumours where data are sparse. Historical control data should be from studies in the same time frame, for the same animal strain, preferably from the same laboratory or the same supplier and preferably reviewed by the same pathologist.^{17 18} While the EFSA final peer review⁴ mentions the use of historical control data from the original laboratory, no specifics are provided and the only referenced historical control data²⁴ are in the BfR addendum.⁵ One of the mouse studies¹² was clearly done before this historical control database was developed, one study (Sugimoto. Unpublished, 1997) used Crj:CD-1 mice rather than Crl:CD-1 mice, and one study¹³ did not specify the substrain and was reported in 1993 (probably started prior to 1988). Hence, only a single study (Unknown. Unpublished, 2009) used the same mouse strain as the cited historical controls, but was reported more than 10 years after the historical control data set was developed.

The RAR dismissed the slightly increased tumour incidences in the studies considered because they occurred “only at dose levels at or above the limit dose/maximum tolerated dose (MTD)”, and because there was a lack of preneoplastic lesions. Exceeding the MTD is demonstrated by an increase in mortality or other serious toxicological findings at the highest dose, not by a slight reduction in body weight. No serious toxicological findings were reported at the highest doses for the mouse studies in the RAR. While some would argue that these high doses could cause cellular disruption (eg, regenerative hyperplasia) leading to cancer, no evidence of this was reported in any study. Finally, a lack of preneoplastic lesions for a significant neoplastic finding is insufficient reason to discard the finding.

Mechanistic information

The BfR Addendum dismisses the IARC WG finding that ‘there is strong evidence that glyphosate causes genotoxicity’ by suggesting that unpublished evidence not seen by the IARC WG was overwhelmingly negative and that, since the reviewed studies were not done under guideline principles, they should get less weight. To maintain transparency, IARC reviews only publicly available data. The use of confidential data submitted to the BfR makes it impossible for any scientist not associated with BfR to review this conclusion. Further weakening their interpretation, the BfR did not include evidence of chromosomal damage from exposed humans or human cells that were highlighted in Tables 4.1 and 4.2 of the IARC Monograph ³

The BfR confirms (p.79) that the studies evaluated by the IARC WG on oxidative stress were predominantly positive but does not agree that this is strong support for an oxidative stress mechanism. They minimise the significance of these findings predominantly because of a lack of positive controls in some studies and because many of the studies used glyphosate formulations and not pure glyphosate. In contrast, the WG concluded that (p.77) ‘Strong evidence exists that glyphosate, AMPA and glyphosate-based formulations can induce oxidative stress’. From a scientific perspective, these types of mechanistic studies play a key role in distinguishing between the effects of mixtures, pure substances and metabolites.

Finally, we strongly disagree that data from studies published in the peer-reviewed literature should automatically receive less weight than guideline studies. Compliance with guidelines and Good Laboratory Practice does not guarantee validity and relevance of the study design, statistical rigour and attention to sources of bias.^{25 26} The majority of research after the initial marketing approval, including epidemiology studies, will be conducted in research laboratories using various models to address specific issues related to toxicity, often with no testing guidelines available. Peer-reviewed and published findings have great value in understanding mechanisms of carcinogenicity and should be given appropriate weight in an evaluation based on study quality, not just on compliance with guideline rules.

General comments

Science moves forward on careful evaluations of data and a rigorous review of findings, interpretations and conclusions. An important aspect of this process is transparency and the ability to question or debate the findings of others. This ensures the validity of the results and provides a strong basis for decisions. Many of the elements of transparency do not exist for the RAR.⁵ For example, citations for almost all references, even those from the open scientific literature, have been redacted. The ability to objectively evaluate the findings of a scientific report requires a complete list of cited supporting evidence. As another example, there are no authors or contributors listed for either document, a requirement for publication in virtually all scientific journals where financial support, conflicts of interest and affiliations of authors are fully disclosed. This is in direct contrast to the IARC WG evaluation listing all authors, all publications and public disclosure of pertinent conflicts of interest prior to the WG meeting.²⁷

Several guidelines have been devised for conducting careful evaluation and analysis of carcinogenicity data, most after consultation with scientists from around the world. Two of the most widely used guidelines in Europe are the OECD guidance on the conduct and design of chronic toxicity and carcinogenicity studies¹⁷ and the European Chemicals Agency Guidance on Commission Regulation (EU) No 286/2011;¹⁸ both are cited in the RAR. The methods used for historical controls and trend analysis are inconsistent with these guidelines.

Owing to the potential public health impact of glyphosate, which is an extensively used pesticide, it is essential that all scientific evidence relating to its possible carcinogenicity is publicly accessible and reviewed transparently in accordance with established scientific criteria.

Summary

The IARC WG concluded that glyphosate is a ‘probable human carcinogen’, putting it into IARC category 2A due to sufficient evidence of carcinogenicity in animals, limited evidence of carcinogenicity in humans and strong evidence for two carcinogenic mechanisms.

• The IARC WG found an association between NHL and glyphosate based on the available human evidence.

• The IARC WG found significant carcinogenic effects in laboratory animals for rare kidney tumours and hemangiosarcoma in two mouse studies and benign tumours in two rat studies.

• The IARC WG concluded that there was strong evidence of genotoxicity and oxidative stress for glyphosate, entirely from publicly available research, including findings of DNA damage in the peripheral blood of exposed humans.

The RAR concluded⁵ (Vol. 1, p.160) that ‘classification and labelling for carcinogenesis is not warranted’ and ‘glyphosate is devoid of genotoxic potential’.

• EFSA⁴ classified the human evidence as ‘very limited’ and then dismissed any association of glyphosate with cancer without clear explanation or justification.

• Ignoring established guidelines cited in their report, EFSA dismissed evidence of renal tumours in three mouse studies, hemangiosarcoma in two mouse studies and malignant lymphoma in two mouse studies. Thus, EFSA incorrectly discarded all findings of glyphosate-induced cancer in animals as chance occurrences.

• EFSA ignored important laboratory and human mechanistic evidence of genotoxicity.

• EFSA confirmed that glyphosate induces oxidative stress but then, having dismissed all other findings of possible carcinogenicity, dismissed this finding on the grounds that oxidative stress alone is not sufficient for carcinogen labelling.

The most appropriate and scientifically based evaluation of the cancers reported in humans and laboratory animals as well as supportive mechanistic data is that glyphosate is a probable human carcinogen. On the basis of this conclusion and in the absence of evidence to the contrary, it is reasonable to conclude that glyphosate formulations should also be considered likely human carcinogens. The CLP Criteria¹⁸ (Table 3.6.1, p.371) allow for a similar classification of Category 1B when there are ‘studies showing limited evidence of carcinogenicity in humans together with limited evidence of carcinogenicity in experimental animals’.

In the RAR, almost no weight is given to studies from the published literature and there is an over-reliance on non-publicly available industry-provided studies using a limited set of assays that define the minimum data necessary for the marketing of a pesticide. The IARC WG evaluation of probably carcinogenic to humans accurately reflects the results of published scientific literature on glyphosate and, on the face of it, unpublished studies to which EFSA refers.

Most of the authors of this commentary previously expressed their concerns to EFSA and others regarding their review of glyphosate²⁸ to which EFSA has published a reply.²⁹ This commentary responds to the EFSA reply.

The views expressed in this editorial are the opinion of the authors and do not imply an endorsement or support for these opinions by any organisations to which they are affiliated.