Helapet Aseptic Study Day 2008

Gene Therapy - Problems and Challenges

Alison M. Beaney Regional Quality Assurance Specialist North-East and Yorkshire

Gene Therapy

Background to Gene Therapy Potential Benefits Perceived Hazards and Risks Regulations Implications for Pharmacy Aseptic Units Future?

Gene Therapy

Definition The deliberate introduction of genetic material into human somatic cells for therapeutic, prophylactic or diagnostic purposes
   

Addition of EXTRA genes Aim is to cure disease (or at least help the patient) First introduction of gene-modified cells into a patient was in 1989 First gene therapy product approved for market in 2004

Still very experimental and early in its development

3

PTQA April 2008

Gene Therapy Vectors

Vectors deliver genes to cells

Therapeutic gene (Transgene)

Transcription

Vector for efficient gene delivery

Translation

Therapeutic protein

Types of Gene Therapy Vectors

Non-viral vectors
  

Naked DNA Liposomes/DNA Polymer/DNA complex (polyplex) Liposome/Polymer/DN A (lipopolyplex)

Viral vectors


DNA viruses
 

Adenovirus Herpes Simplex Virus Retrovirus

6

RNA viruses


Gene Therapy Strategies

1) Gene Replacement
Replace faulty genes with normal genes Corrects inherited genetic errors Provides a missing function Monogenic diseases e.g. cystic fibrosis, haemophilia, X-SCID

2) Gene Addition

Delivers genes to provide a new function Polygenic diseases e.g. cancer

7

2001

Were trying to make a mouse contraceptive vaccine for pest control Used modified mousepox virus as vehicle for transporting antibodies into mice Inserted gene to create IL-4 (interleukin 4) to boost production Surprise !! totally suppressed the "cell-mediated response which combats viral infection

Mousepox 100% lethal

8

December 19, 2007

Boy gets leukaemia after gene treatment to cure bubble baby syndrome

3 year-old with X-linked severe combined immunodeficiency (X-SCID) - immune system fails to develop Treated with genetically modified virus to correct the faulty DNA that causes X-SCID Inserting the replacement DNA activated another gene that promotes cancer Now an acknowledged risk of gene therapy Also seen in 4 / 11 patients in a French trial One has died while 3 are in remission

Retrovirus vector

Regulations governing the handling of gene therapy vectors

No additional regulations governing the handling of Non-Infectious vectors



Non-viral & Non-bacterial

Viral vectors are Genetically Modified Genetically Modified Organisms (GMOs)

10

Genetic Modification

Genetic modification is officially defined as the alteration of genetic material (DNA or RNA) of an organism by means that could not occur naturally through mating and/or recombination
A guide to Genetically modified organisms (Contained Use) Regulations 2000. Health and Safety Executive

11

Regulations governing the handling of gene therapy viral vectors

Two sets of Regulations:


GMO (Contained Use) Regs 2000, HSE



All possible barriers (physical, biological or chemical) are in place to limit contact of the GMOs with humans and the environment All appropriate measures are taken to avoid damage to the environment from the escape or release from human control of GMOs

GMO (Deliberate Release) Regs 2002, DEFRA



aimed at laboratories (difficult to interpret clinically)  no reference to product or patient safety


12

Additional Regulations that apply to Gene Therapy Clinical Trials

Protection of the Patient


Gene Therapy Advisory Committee (GTAC)

Established 1993, Department of Health  UK national research ethics committee (REC) for gene therapy

  

Ethical acceptability for human gene therapy Scientific merits Potential benefits and risks

Patient flagging and long term monitoring  Advice to UK health Ministers on developments in gene therapy research  Applies to ALL GENE THERAPY CLINICAL TRIALS using viral and non-viral vectors 13


Containment Levels for GMOs

Containment Measures Required
Isolatable Lab Suite Microbiological Safety Cabinet Gloves Protective Clothing

Class 1 Level 1 Class 2 Level 2 Class 3 Level 3 Class 4 Level 4

NO NO

NO Risk Assessment R/A YES YES

NO R/A

YES YES

Requires first use of premises notification to HSE Minimum requirement for any human blood or clinical samples. Requires HSE notification Requires HSE notification

YES YES

YES YES

YES
+ Footwear

YES
Complete change of clothing and footwear on entry and exit

Requires HSE notification

Guidelines on Handling GMOs in Pharmacy

QA of Aseptic Preparation Services (4th Edn.)

Appendix 6 Gene Therapy

Scientific Advisory Committee on Genetic Modification (SACGM), Part 6,

Guidance on the use of genetically modified micro-organisms in a clinical setting

European Association of Hospital Pharmacists (EAHP)

Guidance on the Pharmacy Handling of Gene Medicines

Rules and Guidance for Pharmaceutical Manufacturers and Distributors 2007

No Specific Guidance
15

APPENDIX 6 GENE THERAPY

Facilities Documentation Labelling Training Aseptic processing

Cleaning Storage Transport Waste Disposal Spillage

16

Facilities


Gene therapy should not be manipulated in clinical areas

 Basic

Principles - Containment
- Knowledge / understanding /skill - Validated procedures

 

Persons handling the product should be masked and gloved All disposable equipment and materials used for prep & admin - handled as biohazardous

Dedicated facilities required

-ve pressure isolators or Class II BSC  +ve pressure room or lobby  Containment level > 2

17

Clean room suite designed to provide protection to the cleanroom

Aseptic Manipulation


Doses  Calculation/dilutions /multiple dilutions  Needle stick injury risk  Units

 Particle

Units/ml (PU/ml)  Plaque Forming Units/ml (PFU/ml)  Infectious particle Units/ml (IU/ml)  Gene Transfer Units/ml (GTU/ml)


Stability  Container compatibilities - Plastic/glass adhesion  Expiry date - Time to administration from thawing
19

Decontamination

Cleaning


Virucidal detergents (validated against GT vectors)  Cleaning Validation



Specific Detection methods needed for viruses that are virus specific and highly sensitive

Waste Disposal
 On

site validated autoclave for re-usable equipment  Inactivation on-site for Class 3 vectors
Validated autoclave  Incineration  Disinfectant treatment


20

Accidental Exposure

Spillage  Specific to GT vector  Spillage kit

 Contents

( gloves, masks, aprons, goggles, disposable shoe covers, virucidal detergents, absorbent material, disposable forceps & biohazard incineration bag)  Positioned in all GT handling areas
 Notification

to HSE
21

SOPs needed
Safe handling & protection  Storage  Operators


(Not pregnant, breastfeeding or immunosuppressed)

Training  Facilities  Spillage, contamination & needle stick  Waste disposal, cleaning and transport

22

Risk Assessment

Assess each product individually

    

Cytolytic viruses Non-cytolytic viruses Replication competent Replication deficient Class I, II or III

23

What will the Future bring?

  

Dedicated facilities Automation? The first gene medicine in Europe could be licensed in 2008 Licensed closed-system gene therapy products Use of gene therapy as an adjunct to standard therapy e.g. Radiotherapy & Chemotherapy Vector development e.g.
Targeted vectors (viral & non-viral)  Bacterial vectors

24

 

Additional Information
       Gene Therapy Advisory Committee (GTAC) http://www.advisorybodies.doh.gov.uk/genetics/gtac/index.htm Gene therapy trials worldwide. Provided by the Journal of gene medicine http://82.182.180.141/trials/index.html A guide to Genetically modified organisms (Contained Use) regulations 2000. Health and Safety Executive Genetically Modified Organism (Deliberate Release) Regulations 2002 [GMO(DR)]. Department for the Environment, Food and Rural Affairs (DEFRA) http://www.opsi.gov.uk/si/si2002/uksi_20022443_en.pdf Quality Assurance of Aseptic Preparation Services Fourth Edition. A.M. Beaney. Pharmaceutical Press 2006. Appendix 6. Gene Therapy. EU Clinical Trials Directive. http://www.wctn.org.uk/downloads/EU_Directive/Directive.pdf Implications of gene therapy for hospital pharmacists. Simpson.J, Stoner. N. www.pjonline.com/pdf/articles/ pj_20030726_genetherapy.pdf
25

Additional Information
Simpson. J, James. J.D. Drug Delivery Systems and Sciences 2002, 2 (1), 513.  Standards for gene therapy clinical trials based on pro-active risk assessment in a London NHS Teaching Hospital Trust. Bamford, K.B., Wood, S., Shaw, R.J. QJM 2005, 98, 75-86. www.qjmed.oupjournals.org  Progress in Gene Therapy are hospital pharmacies the next barrier? Simpson, J. Hospital Pharmacist, 2006, 13 (8), 266 http://www.pjonline.com/pdf/hp/200609/hp_200609_comment.pdf  Cancer Biotherapy. An Introductory guide. Young, A. Rowett, L. Kerr, D. Oxford University Press 2006 Scientific Advisory Committee on Genetic Modification (SACGM), Part 6, Guidance on the use of genetically modified microorganisms in a clinical setting. http://www.hse.gov.uk/biosafety/gmo/acgm/acgmcomp/part6.pdf European Association of Hospital Pharmacists (EAHP) Guidance on the Pharmacy Handling of Gene Medicines. http://www.ejhp.eu/

 Cancer gene therapy: from science to clinical trials. Searle. P.F, Spiers. I,

26