golden rice update - specialtycropassistance.org...challenges faced by public and small developers....

DONALD J. MACKENZIE, PH.D.INSTITUTE FOR INTERNATIONAL CROP IMPROVEMENTDONALD DANFORTH PLANT SCIENCE CENTERS C R A W O R K S H O P

4 O C T O B E R 2 0 1 8

The Golden Rice StoryRealizing the Promise of Public

Sector Plant Biotech Product Delivery

More than two decades of work aimed at delivering public-good plant biotech solutions in the developing world

Tens of millions $ invested

Promising technical solutions, but few examples of these entering the market

Lack of institutional experience in product development within a regulated environment

Research projects vs. product delivery

WHERE ARE THE HUMANITARIAN AGRI-BIOTECH PRODUCTS?

Challenges Faced by Public and Small Developers

Limited financial resources to undertake long-term development efforts (typical grant durations 25-30% of product development cycle)

Limited in-house regulatory affairs expertise or experience

Typically, the sole advocate with regulatory agencies on issues related to risk assessment and regulatory practice

Lack specialized organizational structure (regulatory science, quality assurance, product stewardship, etc)

Lack institutional capacity or opportunity to retain and build upon lessons learned each time a product moves through the pipeline

Regulatory data constraints

Unlikely to fully account for the ongoing duty of care to maintain product authorizations

Limited experience in product launch planning, post-release product stewardship, and liability issues in case of AP or LLP

To Add to the Complexity…

Many of the countries where we want to have impact have –

◦ a lack of well-developed product regulatory pathways◦ absence of clear guidance on the regulatory studies

required to demonstrate safety◦ weaknesses in existing risk assessment, inspection, and

enforcement capability◦ limited capability for identity preserved production

practices (commodity streaming)◦ porous borders, informal seed systems

Our products may be the first “locally developed” biotech products undergoing regulatory assessment

Need for capacity building

Empowering our national partners to have “ownership” is essential

Diversity of crop species

Pest and disease resistance, micronutrient biofortification, and abiotic stress tolerance

Target countries have generally weak or newly evolving regulatory systems

Development institutions typically working on their first regulated product

Have the resources to undertake studies that are needed but not studies that are optional (nice to know)

Regulatory harmonization is not optional, it is a must-do for these projects to be successful

The ProblemMany people in developing countries cannot access or afford a diverse and healthy diet

Rice often makes up to 70% or more of daily caloric intake in Bangladesh

However, it is mainly carbohydrates with some protein, but deficient in micronutrients

Micronutrient malnutrition or “hidden hunger” is a serious problem

More than 2 billion people globally are affected by micronutrient deficiencies (iron, zinc, vitamin A)

PREVALENCE OF SUBCLINICAL VAD IN BANGLADESH

Population Group N Prevalence (%)95% Confidence

IntervalPreschool Age Children (6-59 months)

National 873 20.5 [15.9, 25.0]Rural 306 19.4* [13.6, 25.2]

Urban 305 21.2**** [15.5, 26.8]Slum 262 38.1**** [28.4, 47.7]

School Age Children (6-14 years)National 1266 20.8 [17.3, 24.4]

Rural 431 20.1 [16.2, 24.0]Urban 431 22.2*** [12.8, 31.5]

Slum 404 27.1 [21.5, 32.6]*Rural v. urban, P=0·01; **urban v. slum, P=0·003; ***urban v. slum, P=0·004; ****rural v. urban, P=0·006; *****slum v. rural, P<0·001; ******urban v. slum, P<0·001.

Approaches to Combating VADNutrition education and consuming diverse and nutrient rich diet

Promoting breastfeeding and complementary feeding practices

Vitamin A capsule supplementation

Food fortification

Other public health measures, including control of infectious diseases

These approaches have had real successes, however millions are still suffering from VAD

Target populations are sometimes missed with these interventions, especially the hard to reach areas

OUR MISSION | IMPROVE THE HUMAN CONDITION THROUGH PLANT SCIENCE

~18 µg/g β-carotene equivalents

~0.1 µg/g β-carotene equivalents

~1 µg/g β-carotene equivalents

Product of conventional breeding and selection

PROVITAMIN A BIOFORTIFIED MAIZE – NO SAFETY CONCERNS

Provitamin A Biofortified RiceNo Safety ConcernsProduced using biotechnology

Expresses three new enzymes

Phytoene synthase from maize

Phytoene desaturase from bacteria

Phosphomannose isomerase from E. coli for selection in tissue culture

Has been subject to comprehensive safety assessment

No β-carotene

~12 µg/g total carotenoids(GR2E BRRI dhan29 following 2 months storage)

Golden Rice – an illustrative example

• 35 months

• Generation of all regulatory safety data

• Regulatory applications submitted in five jurisdictions

• Three approvals secured for use in food, including completion of FDA biotechnology consultation

21 3

Considerations for Strategy Development

Philippines◦ Despite dozens of prior import approvals for GM

foods/feeds, there has never been an authorization for something not already reviewed by US FDA or another regulatory authority – never been first

◦ Court challenge raised around Bt eggplant and subsequent SC decision resulted in massive disruption of regulatory processes

Bangladesh◦ Limited, conditional release of Bt eggplant in the fall of

2013◦ Unpredictable and non-transparent regulatory processes◦ Poor precedents set by previously submitted regulatory

studies

In both countries, regulatory officials and agencies will be under a high degree of scrutiny over any Golden Rice application

The safety assessment of Golden Rice must be exemplary


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There have been more than 1,260 food safety decisions across 28 countries/regions on biotech crops – never with a differing opinion ILSI Research Foundation, GM Crop Database, March 2017

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# of Approvals # of Events

Elements of the Regulatory Strategy

Focus beyond just the intended countries of cultivation –secure authorizations (food/feed) with other well-respected agencies

◦ Validation of our data package◦ Confidence building within regulatory community◦ Risk mitigation (AP/LLP)

Meet internationally accepted norms/standards, where they exist

Encourage cooperation between regulatory agencies during the review process

Utilize credentialed/qualified CROs – quality analyses and products, on time and budget

Global dossier (food/feed) – easily adjusted to meet local administrative requirements, but same core data package

Transparency and pubic disclosure permitted for all studies and submissions


• Southern blots to demonstrate copy number and inheritance of inserted DNA across generations (use same generations for trait inheritance)

• Sequencing to confirm insert organization and flanking regions – allergen & toxin DB searches if any junctional ORFS (start-to-stop, 30+ aa)

• Insert segregation within 3 segregating generations measure using zygosity PCR

• Produce CRTI and ZmPSY1 test proteins (outsource), fully characterized, active

• SGF digest (No SIF), heat inactivation, seqsimilarity searches against allergens & toxins (E = 1 x 10-5)

• Acute oral toxicity for CRTI only – but not at limit dose (i.e., not 2000 mg/kg), HOSU for ZmPSY1, LOA for PMI data

• Outsource oral tox study to Stine Haskell• No equivalence studies with plant-expressed

CRTI, ZmPSY1, PMI – western blots to compare plant and microbial expressed

• Comp, protein expression, agronomics – collect from 4 sites x 2 seasons

• Measure all comp analytes recommended in the OECD consensus document for rice (grain and straw samples)

• No whole food feeding study (90-day rat or livestock animal)

• No NTO studies

CORE PROJECT STUDY PLAN


Studies submitted to Food Standards Australia New Zealand, Health Canada, US Food and Drug Administration, Philippines Bureau of Plant Industry, Bangladesh

Every regulatory agency received exactly the same data set

All of these studies are publicly available

Molecular Characterization Conclusions

One copy of the pSYN12424 T-DNA was introduced at a single site within the rice genome (chromosome 3)

The introduced DNA and phenotypic trait were stably inherited over multiple generations

The introduced DNA segregated as a single locus according to Mendelian rules of inheritance

The T-DNA was introduced without modifications, deletions, or rearrangements except for small truncations at the 5’ and 3’ termini of 23 bp and 11 bp, respectively

There were no new novel open reading frames created as a consequence of the insertion that would have the potential to encode a protein with any significant amino acid sequence similarity to known toxins or allergens

ZmPSY1 Safety

History of safe use based on known exposure from maize consumption

Mode of action of ZmPSY1 is not related to modes of action of known toxins

No significant amino acid sequence similarity to known allergens or toxins

Rapidly digested within 30 seconds in the presence of simulated gastric fluid containing pepsin and will thus have the same metabolic fate as any other dietary protein Rapidly and irreversibly inactivated following brief heating to 50ºC, a temperature significantly lower than cooking temperatureVery low concentrations in mature grain – 245 ppb (0.00034% of total protein)

CRTI Safety

Mode of action of CRTI is not related to modes of action of known toxins

No significant amino acid sequence similarity to known allergens or toxins

Rapidly digested within 30 seconds in the presence of simulated gastric fluid containing pepsin and will thus have the same metabolic fate as any other dietary protein

Rapidly and irreversibly inactivated following brief heating to 55ºC, a temperature significantly lower than cooking temperature

Demonstrated lack of acute oral toxicity in mice at a dose of 100 mg/kg body weight, representing a margin of exposure of 115000-fold relative to any realistically conceivable human dietary exposure

Very low concentrations in mature grain – 30 ppb (0.00004% of total protein)

PMI Safety

Known function of PMI in catalyzing the reversible interconversion of mannose-6-phosphate and fructose-6-phosphate is ubiquitous in nature and not similar to the activities of known protein toxinsNo significant amino acid sequence similarity to known toxins or meaningful similarity to known allergens

Rapidly digested within 1 minute in the presence of simulated gastric fluid containing pepsin and will thus have the same metabolic fate as any other dietary proteinRapidly and irreversibly inactivated following brief heating to 65ºC, a temperature significantly lower than cooking temperature

Lack of acute oral toxicity in mice at a dosage of 2000 mg/kg body weight, representing a margin of exposure of 69000-fold relative to any realistically conceivable human dietary exposureHighest concentration in mature grain: 1.89 μg/g – 0.0027% of total protein

Safety of PMI has been assessed by regulators in at least 21 countries


• 2015 – Evaluation of 170 BC5F3 introgression lines in BRRI dhan29 in the screen house at Gazipur

• 2016 boro season – CFT evaluation of 23 selected BC5F4 introgression lines in BRRI dhan29 at Gazipur

• 2017 boro season – CFT evaluation of 8 selected BC5F5 introgression lines in BRRI dhan29 at 5 locations

• 2018 boro season – CFT evaluation of 1 BC5F6 introgression line in BRRI dhan29 at 5 locations to advance for seed production

• Unconfined environmental release application submitted by BRRI on 26 November 2017

“No potential public health and safety concerns have been identified in the assessment of GR2E. On the basis of the data provided in the present Application, and other available information, food derived from GR2E is considered to be as safe for human consumption as food derived from conventional rice varieties.”

Approved: 20 December 2017http://www.foodstandards.gov.au/code/applications/Pages/A1138GMriceGR2E.aspx

• “After reviewing the accomplished biosafety forms and socio-economic, ethical and cultural questionnaire, and supporting documents submitted by the applicant, the assessors namely: Scientific and Technical Review Panel (STRP), BPI- Plant Products Safety Services Division (BPI-PPSSD) and Bureau of Animal Industry (BAI), concurred that GR2E Rice is as safe for human food and animal feed as its conventional counterpart.”

GM golden rice gets approval from food regulators in the US 24 May 2018

Looking Ahead for Golden Rice in

BangladeshBiosafety review is now with the Biosafety Core Committee under DOE

Pending recommendation, next step is endorsement by NCB

Farmer and consumer quality testing

Completion of proposed variety trials in 2019 and submission of data to the National Seed Board for variety recommendation

Potential release of the first Golden Rice varieties at the end of the decade


Mutual Recognition of Safety Assessments

Mechanism for gaining risk assessment resource utilization efficiency – utilizing another authority’s

work product for decision-making

Can drive greater consistency in dossier submissions – one core dossier can be used for multiple

jurisdictions

Promotes greater synchrony in submission timing between jurisdictions, and possibly decision-making

Is the only practical solution to addressing issues related to LLP and trade disruption

• Regulatory success is achievable for public / small developers• 3Ds – discipline, diligence, and determination – essential to

success• Sound science is fundamental to regulatory harmonization• Regulatory harmonization does not compromise national

autonomy but requires political will• It does promote science-based decision-making• Significant benefits will accrue to the public sector and SMEs

Thank You!

Donald J. MacKenzie, Ph.D.Executive DirectorInstitute for International Crop ImprovementDonald Danforth Plant Science Center975 North Warson Road, Saint Louis, MO 63132Office: +1 (314) 587-1612 Fax: +1 (314) 587-1712 Cell: +1 (202) 695-0436

golden rice update - specialtycropassistance.org...challenges faced by public and small developers....

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