Environmental assessments and shedding studies
for gene therapy products in the US and in the EU

Kanazawa Institute of Technology
Teruhide Yamaguchi

2018.11.12
The 15th DIA Japan Annual MeetingEnvironmental Assessment for Gene Therapy
Products and Virus-shedding Study<Health Care
professional>

Virus Vector

Viral Shedding???
secretion ・ Emissions(Urea, faeces, sputum）???

???

Administration<Family><Environment>

Environmental assessment and guideline for viral sheddingin USA USA does not ratifiy the Cartagenus law
 According to US federal law, the sponsor should submit theenvironmental assessment about gene therapy product.
 The National Environmental Policy Act of 1969 (NEPA);
Revision of Policies and Procedures (Final Rule) 1997
 federal law 21CFR25

 Determining the Need for and Content of Environmental
Assessments for Gene Therapies, Vectored Vaccines, andRelated Recombinant Viral or Microbial Products（Guidance for Industry, FDA CBER, March 2015）
 Design and analysis of shedding studies for virus or bacteriabased gene therapy and oncolytic products（Guidance for Industry, FDA CBER, 2015)

FDA Guideline for the Environmental
Assessment: IND Approval The sponsor should submit the following data in IND（21CFR25.15(a)）
 Environmental assessment
 Claim for categorical exclusion
 In the case of no categorical exclusion：
Extraordinary circumstances indicate that the specific proposed agency
action may significantly affect the quality of the environment
 Extraordinary circumstances ：Specific or unknown risk on human 、Risk
on Endangered Species of Wild Fauna and Flora (CITES) In most cases, IND for a clinical study using a GTVV will notsignificantly affect the quality of the environment because, in brief,these clinical trials are closely monitored and are limited to a
designated study group.

FDA Review of Environmental Assessment for INDof GMO products
 Key Issues of Environmental Assessment for Human Gene Therapy
Investigational New Drug Applications (INDs).
 Collection of viral shedding data to evaluate the environmental
assessment
 Measures to mitigate the potential risk for securing the safety of theinvestigational study
 Environment Reviewer is responsible for evaluation of the environmentalrisk)

 Emergency of replication competent virus is concern risk for replication
incompetent virus vector including recombinant viral vaccine
 Recommend to collect the viral shedding data about 10 number ofpatients➡If the viral shedding risk is low, it may be skip to collect the data of viralshedding.
 If the data or monitoring protocol is not enough, FDA submit the clinicalhold to the sponsor

FDAのEnvironmental Assessment of Human
Drug and Biologics Applications：（BLA) BLA of gene therapy products：the sponsor should submit EA
 categorical exclusion （Substances that Occur Naturally in the Environment）are; contain functional protein-coding sequences from one or more species within asingle genus to “occur naturally in the environment” for purposes of 21 CFR
25.31(c).

 differ from a wild-type substance only in attenuating point
mutations or deletions to be substances that “occur naturally inthe environment” for purposes of 21 CFR 25.31(c) have been killed or inactivated by undergoing a specificmanufacturing step designed to eliminate their ability to replicate
to be substances that “occur naturally in the environment”
 consist of genetically-modified human cells to be substances that“occur naturally in the environment”

FDAのEnvironmental Assessment of Human
Drug and Biologics Applications：（BLA) BLA of gene therapy products：the sponsor should submit EA

 Identification of Substances Subject to the Proposed Action“Identification of Substances Subject to Proposed Action”; “Identifyingand Assessing Potential Environmental Effects”; “Mitigation Measures”;
and “Alternatives to the Proposed Action.” Identification of Substances Subject to the Proposed Action
To identify known and potential variants of the GTVVs released intothe environment.
 Identifying and Assessing Potential Environmental Effects
 Is the vector virulent, pathogenic, What is known regarding thegenetic stability and prevalence of gene exchange in natural
populations of the strain or vector?
 the product have traits that may give it a selective advantage overnatural organisms?
 Assessing the magnitude of potential environmental effects
 What is known about the effectiveness of monitoring and mitigation plans?

FDA Environmental Assessment for GMO products
 The release of vector DNA into the environment is detectable by PCR
at the injection site and/or in excreta Replication competent impurity may cause an active infection in thepatient or study participant capable of disseminating to others. ⇒These variants may arise during manufacture or after productadministration because of mutations, or recombination events.
 A super-coiled plasmid may still retain the ability to transfer genetic
material (such as antibiotic resistance genes) to other bacteria evenafter limited degradation.

Levy et al: Quantitation of supercoiled circular content in plasmid DNA solution
using fluorescence based method. NAR 28, e57, 2000

FDA: Design and Analysis of Shedding Studies for Virus or BacteriaBased Gene Therapy and Oncolytic Products
1. Why this guideline is published?
-> The possibility that the shed viral vector for gene therapy or oncolytic product may be infectious
raises safety concerns related to the risk of transmission to untreated individuals..
-> There are many product-specific factors and patient-specific factors that can influence the design ofa shedding study.
2. Purpose of the viral shedding guideline ？
->Recommendations on how to conduct shedding studies during preclinical and clinical development
How and when shedding data should be collected ？
-> How shedding data can be used to assess the potential for transmission to untreated individuals.
3. What is shedding-> Excreta (feces); secreta (urine, saliva, etc); or through the skin (pustules, sores, wounds)-> Shedding is distinct from biodistribution, and signal in blood is not shedding
4. Design of Shedding Studies: Guiding Principles？
-> Choice of samples, Frequency of sample collection, Duration of the monitoring period, Assay-> VBGT Characteristics（Replication competence, Immunogenicity, Tropism) Route of Administration
・ Replication competence：Start from P-1 to P-2, Assess before P-3、・ Replication Incompetence：Start after the determination of dosing (P-2)
・ Examples：sampling on the day, 1、3、7、10day then weekly. Continue to below LOD

・ Samples should be determined in design of shedding studies
・ Replication competence：Infectivity Assay + PCR、incompetent：PCR

PCR (+) -> Infectivity (?)

According to shedding data, the sponsor should assess the risk of transmission to untreated individuals

EU Environmental Assessment: Law and Guideline
 EU and European nations have ratified the Cartagena Law
 EMA reviews the environmental risk of gene therapy product insubmission of new drug approval centrally; Evironmental Risk
Assessment + Control of Viral Shedding Directive 2001/83/EC, as amended, and Regulation
726/2004
 Guideline on Scientific Requirements for The Environment
Risk Assessment of Gene Therapy Medicinal Products(EMEA/CHMP/GTWP/125491/2006)
 ICH Considerations: general principles to address virus
and vector sheddingEU: Environmental Assess of GMO for IND
 For products containing genetically modified organisms (GMOs), an additional risk
assessment step in the clinical trial authorisation (CTA) procedure is required inaddition of IND.
 The application process for conducting clinical trials with an ATIMP consisting of orcontaining a GMO in the EU involves the need for differentiation between contained
use and deliberate release review by different responsible authorities, with specificdocuments and procedures in addition to the standard CTA review by a competent
health authority and the Ethics Committee. ➡ A multi-stakeholder meeting at EMA
regarding the lack of harmonisation of the GMO requirements and the challenge tointegrate the GMO assessment in the clinical trial process.
 The focus of Directive 2009/41/EC is on the assessment of the biosafety levelclassification of the GMO and the implementation of physical, chemical andbiological barriers. All contained use procedures are classified based on theperformed risk assessment into one of the following four categories:

Category

Risk Assessment

Viral Vector

Class 1

No or negligible risk

AAV（helper free)Class 2

Low risk, level 2 containment

AAV (AdV helper), AdV, HSVClass 3Moderate risk, level 3 containment

Lentivirus

Class 4High risk, level 4 containment

GMO Contained use vs. Deliberate release
GMO –Contained use•

Contained use is defined as any activity with GMOs for which specific containment
measures are used to limit their contact with the environment.

•The focus of Directive 2009/41/EC is on the assessment of the biosafety levelclassification of the GMO and the implementation of physical, chemical and biologicalbarriers. The risk classification has consequences for the procedure and review periodof the application
Contained use –risk classification
All contained use procedures are based on classification of risk –as decided byappropriate agency / authority
Class 1: No or negligible risk, level 1containment
Class 2: Low risk, level 2containment
Class 3: Moderate risk, level 3containment
Class 4: High risk, level 4containment
Classification is dependent upon strength of argument / data contained in a risk
assessment / biosafety dossierClass 3 / 4 GMOs require prior consent from the competent authority but most ATMPs
should be Class 1 / 2
Contained use often requires clinical site-specific notifications and / or submissions toauthorities••••

•
•
•
•

GMO -Deliberate release
Deliberate release is defined as any activity with GMOs that is not contained use.
Directive 2001/18/EC is based on a environmental risk assessment (ERA) coveringeffects on human health or the environment.
The ERA should be carried out in accordance with the principles set out in Annex IIof this Directive. 5 steps:i) identification of potential adverse effects,ii) estimation of the likelihood,iii) risk estimation,iv) risk managementv) assessment of the overall environmental impact.

Contained use vs deliberate release
Generally, GMOs regulated under deliberate release come under greater scrutiny fromcompetent authorities
Contained use has a greater administrative burden (requiring both clinical sitesubmissions and sponsor submissions) which may slow approvals
However, lower classification equates to lighter regulatory touch and once approved,
there are less likely to be conditions applied
Deliberate release or high risk contained use may require significant safety data / risk
mitigations prior to or during the trial

Clinical trials using GMOs classified as contained use
or deliberate release for selected EU member states
MemberState
Germany

Contained use or Deliberate release

UKEither (most studies are considered contained use)

FranceEither(until recently all studies considered
contained use but deliberate release alsoapplicable)
Clinical studies are now normally considered as
deliberate release
EitherSwedenSpain

Deliberate release

Deliberate release
TheNetherlands
Belgium
Either

Gene Therapy Legislation in Germany
Gene therapy trials require authorization by the Paul-Ehrlich-Institut (www.pei.de), residing
with the Ministry of Health (Bundesministerium für Gesundheit), and approval by the local
ethics committee of the principal investigator. Guidance on the submission of clinical trial
applications is provided by the 3rd Notification on the Clinical Trial of Medicinal Products forhuman Use. Germany generally considers clinical trials with medicinal products containing
genetically modified organisms (GMOs) as deliberate release and requires an environmental
risk assessment (ERA) in accordance with Annex II and based on information as per Annex III
to the Directive 2001/18/EC.
The deliberate release of a GMO within a clinical trial is authorised by the Paul-EhrlichInstitut in consultation with the German GMO competent authority, the Federal Office ofConsumer Protection and Food Safety (BVL). Any handling of the GMO-containing medicinal
product outside the clinical trial falls within contained use which is regulated by the Genetic
Engineering Act (GenTG).

Database are published in GMOinfo-GMOregisterof European Commission

SUMMARY NOTIFICATION INFORMATION FORMAT FOR THE RELEASE OF GENETICALLY MODIFIED ORGANISMS
OTHER THAN HIGHER PLANTS IN ACCORDANCE WITH ARTICLE 11 OF DIRECTIVE 2001/18/EC

Title of the project: Oncolytic adenovirus therapy using a prostate-specific conditionallyreplication-competent adenovirus as an adjuvant treatment for localized prostate cancer.
GMO named: Genus Mastadenovirus, species human Adenovirus subgenus C,
serotype 5.

Summary of the potential environmental impact of the release of the GMOs

The environmental risks associated with the use of the GMO are negligible. The GMO isa strongly attenuated derivative from a commonly occurring cold virus with anunmodified tropism, and adverse effects are limited to the prostate only. There is a riskof transmission of this virus in case of an accident during administration, resulting in thepotential exposure of pharmacy or trial personnel to high amounts of virus, and in caseof exposure of third persons by transmission of virus that is shed via excreta,
predominantly semen. Harmful effects of transmission of virus or recombinants to thirdpersons are not to be expected due to the strongly attenuated pathogenicity of theGMO, which is restricted to prostate cells only, and the negligible level of potentialrecombinants.
Identity of the vector,
Host range of the vector] Description of identification and detection methods(a) Techniques used to detect the GMO in the environment(b) Techniques used to identify the GMO
4. Method and amount of release(a) Quantities of GMOs to be released:(b) (c) Methods and procedures to avoid and/or minimise the spread of the GMOs beyond the siteSummary of GMO regulation in EU Nations•••••

GMO requirements are implemented very differently across Europe and canrequire regional interactions with numerous bodies / authorities
Admin burden can be great for small companies and lack of clarity withprocedures can be frustrating
Benefits of using contained use procedures depends upon MS but generally
seen to be less burdensome in terms of negating risk to humans andenvironment
Deliberate release or higher risk contained use (Class 3/4) may requiresignificant safety data / risk mitigations prior to, or during the trial
Its not about contained use vs deliberate release –just need the approval!

UKにおけるContained Useの考え方
 CU：GMOが環境中に放出される可能性がほとんどない場合
 物理的バリア（入院管理）
 生物的バリア（十分弱毒化されている場合、感染性、増殖性、外界での生存性を失っている場合）
 Sheddingがほとんどない場合

 入院管理はCUだが、入院しなければDRとは限らない ワクシニアワクチンでは密封包帯を使用すればCU
 長期間排出が予測される場合はDRとして扱い、sheddingのモニタリングを実施
CUの場合、治験前の環境影響評価は不要で承認申請時に実施

EMA 審査報告書
Strimvelis (ADA導入自己CD34細胞)
環境毒性＆環境リスク評価
 対象は第三者及び動植物を含む環境全般
 評価結果は非臨床の毒性の項に記載
 遺伝子導入された細胞（Strimvelis)とADA retrovirusの特性を考慮
 医療従事者への影響を避けるため治療場所を限定した使用 投与部位からの漏出を防ぐ非開放系での使用により環境への伝播を防止
 20年近くにわたる使用経験から環境へのリスクが少ないことを確認

 増殖性ウイルス（RCR)の出現リスクについても考察：➡臨床試験でRCRの検出がないこと、患者体内で野生型のレトロウイルスと組換えが起こりにくい設計となっている（ホモロジーをできる限り低減化）

EMA 審査報告書
Zalmoxis (HSV-TK導入同種T細胞)
環境毒性＆環境リスク評価
 対象は第三者、動植物を含む環境全般
 評価結果は非臨床の毒性の項に記載
 遺伝子導入された細胞（Zalmoxis)とγ-retrovirusの特性を考慮したリスク評価
 医療従事者への影響を避けるため治療場所を限定した使用 Phase I/IIとPhase IIIで2つのベクターを使用
 ベクターの伝播リスクは、ベクターそのもの又はプロウイルスが生殖細胞に入ったときのみ起こりうると推定
 Viral-sheddingに関する解析は実施されていない（遺伝子組換えT細胞にはフリーのウイルスベクターは含まれていない）
 生殖細胞へのベクター分布のリスクはないと判断（3箇所の変異を導入したベクターのためRCR出現のリスクはほぼない、T細胞系のみ増幅する条件etc.）

EMA 審査報告書
Imlygic (腫瘍溶解性ウイルス)
環境毒性＆環境リスク評価

 遺伝子組換え制限増殖性ウイルス（HSV-1由来）の開放系での使用
 神経毒性や病原性は1/100～1/1000に減弱されており、また、がん細胞でのみ増幅し正常細胞では増幅しない設計（ベクターの安全設計）

 最も伝播の起こりやすい第三者は医療従事者と近親者と想定
 投与部位からの排出が最もハイリスク
 対象疾患を成人のメラノーマに限定することにより、環境中への排出によるリスクを低減化

 Imlygicは患者から排出されると急速に感染性を失うことから、環境影響リスクは少ないと考えられる

 しかし、免疫抑制状態にある患者や妊婦へ暴露された場合のリスクは健常人に比べて高いと考えられる

Environmental Risk of Replication Competent Virus and ViralVector
 Recombinant Oncolytic Virus Vector
 Wild and Attenuated Oncolytic Virus Attenuated Viral Vaccine expressing recombinant Antigen (Ebola,MERS antigen etc.)
 Imlygic, recombinant oncolytic HSV-1 virus has a risk on immunecompromised patients, new-born children.
 The sponsor was required to mitigate the risk on transmission of Imlygic
to intimate 3rd party and HCW.
 To prevent the 3rd party transmission, the sponsor provides themedication-guide and the education program to patients
Sponsor is required to prevent the transmission of vectors from patients to3rd parties in clinical use of gene therapy products (ICH Consideration, viralshedding).
免疫状態の正常なヒトに対しては安全性上問題はなくても、免疫不全の人に対しては重篤な副作用を及ぼす可能性がある。そのリスクを低減する方策が必要。

Comparison of GMO Regulation about Gene Therapy
Products between EU, USA and Japan
JapanUSAEU

Cartagena Law

Adapt

No

Adapt

Environment
Assessment GLNone

Adapt

Adapt

Focus

Recombinant Virus

Gene Therapy Products
include PlasmidAt Approval for Market
Authrization (MA)(except for replicationcompetent virus or etc）

Gene Therapy Products
include Plasmid

Review Timing

Before Clinical Study

Viral Shedding

ICH Consideration

Before Clinical Study byeach Regulatory Agency
At Approval for MA（EMA）

Viral Shedding GL

ICH Considration

Shedding Data

Submit with Approval for MA

Submit with Approval for MA

When collectshedding data

Replication Competent Virus:
Phase I
Non-replication : Phase II

England：Phase I

In or out hospital

In or out hospital

Control Pts

Phase 1:
In hospital (isolatedroom)*Cartagena law

 Japanese Cartagena Law may correspond EU and USA environmental Assessment.

遺伝子治療薬開発ステージと環境影響評価、カルタヘナ申請
R&D

治験初期に緻密なデータ収集

米国

Consultation

EU

基礎
研
究

非臨床試験

Pre-Pre IND
meeting

Pre IND
meeting

非臨床試験

各国でのGMO規制
国による審査対応の違い

Approval forMarketAuthrization

治験届

第2種使用
第1種使用

基礎
研
究

MarketPh-III

Non-clinical

RS consul.

Ph-II

Ph-I

CMC

日本

Clinical Trial

IND（第治一験相）

（前治期験
第二相）

（後治期験第二相）

承認時の評価と承認後の使用条件？
（第治三験相）IND（第治一験相）viral shedding
Dataの収集

申請と承認RMPEA治験でのViral
sheddingデータ

viral shedding
Dataの収集

（前治期験
第二相）

RMP

（後治期験
第二相）

（第治三験相）

申
請と承認

RMP

ERA治験でのViralsheddingデータ

SummaryThank you !
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