Quality and GLP

for Histology and Pathology of

Drug Safety Studies

Roger Alison BVSc MRCVS DiplECVP

Consultant Toxicological Pathologist

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What is Quality Histology?

It depends upon the purpose

- Answer research questions

- Diagnosis

- Screening

- Support regulatory submissions

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Quality Regulatory Histology

Fitness for the purpose

- Acceptability

- Reliability

- Consistency

- Reproducibility

- Retrievability

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Quality Systems

- Good Laboratory Practice (GLP)

- Others:

- Good Clinical Practice (GCP)

- Good Manufacturing Practice (GMP)

- ISO 9000

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GLP

Definition

- GLP is a quality system concerned with

the organisational process and the

conditions under which non-clinical

health and environmental safety studies

are planned, performed, monitored,

recorded, reported and archived.

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Purpose of GLP

- Ensure consistent, reliable standards of

safety testing world-wide

- Promote the quality and integrity of test

data

- Improve human and environmental

safety

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Origin of GLP

Introduced because of fraud or poor quality

work in US in 1970s:

- Dead animals “reappeared”

- Tumours “disappeared”

- Inadequate mixing of dose in diets

- Inadequate/insufficient documentation

- Replacement of animals

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GLP is necessary...

Ethically: prevent fraud, public risk,

unnecessary use of animals

Economically: avoid waste of time and money

Scientifically: avoid acceptance of false

results

Regulatorily: ensure confidence in regulatory

decisions

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GLP is required for...

All studies to assess the safety of:

- Drugs and medical devices

- Pesticides

- Chemicals

- Biologicals

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Personnel

Sponsor

- Study Monitor

Test Facility/Test Site(s)

- Management

- Study Director/Principal Investigator(s)

- Study Personnel

- Lead Quality Assurance (facility)

- Test Site Quality Assurance Unit(s)

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Sponsor

Commissions and submits GLP work

Responsible for:

- Ensuring laboratory is GLP compliant

- Test material information

- Approval of protocol

- Archiving – if contract laboratory or test

facility goes out of business

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Management

Ensure GLP complied with:

- Designate a Study Director

- Ensure staff are trained and experienced

- Ensure adequate resources are available

- Approve SOPs

- Ensure QA programme exists (and

reports to management)

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Study Director

The single point of study control

Takes responsibility for the overall conduct of

the study and its final report

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Study Director ensures...

- GLP is followed

- Adequate trained personnel and suitable

equipment

- Personnel approve the protocol

- Protocol, amendments and SOPs are

available to study personnel

- Procedures specified in the protocol are

followed

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Study Director ensures...

- Unforseen events are noted and assessed

- Raw data fully documented

- Approves and signs the report

- Archives study material promptly on

completion of the study

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Principal Investigator

At sites separated from the SD, the PI

oversees critical phases of the study

- Drafts relevant protocol sections

- Ensures work is conducted to GLP and

in accordance with protocol and SOPs

- Prepares contributing report

- Ensures raw data is archived

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Study Personnel

- Know Principles of GLP applicable to

their involvement in the study

- Ensure their training record is up to date

- Follow protocols, SOPs

- Report problems, mistakes, unexpected

events, and involved in corrective action

- Record data promptly, accurately,

completely

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Quality Assurance

Independent of study conduct

- inspect study at intervals

- protocol

- in-life data, procedures

- report

- facilities

- Report to management and SD

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Protocols and SOPs

Specify how a study is to be conducted:

- What is going to be done

- Who will do it

- When will it be done

- What data will be recorded

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Protocols and SOPs

Protocols (study plans)

- Study specific instructions, often linking

SOPs and providing timings

SOPs:

- Standard: routine

- Operating: working

- Procedures: methods

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Protocol

GLP requires:

- A written plan, before start of the study

- Verified for compliance by QA

- Approved by test facility management

(and usually by sponsor)

- Approved by dated signature of Study

Director

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Protocol

Informs:

- Study staff, support staff

- Management

- Subcontractors, Principal Investigators

Confirms responsibilities (by signature):

- Study Director

- Management

- Principal Investigators

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Protocol

Information included:

- When animals to be killed

- Tissues to be taken at necropsy

- Tissues to be processed

- Slides made: group, numbers, staining

- Slides examined (control, high dose?)

- Special techniques

- Staff responsible

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Protocol Deviations

Protocol Amendments: (study plans)

- Planned changes to the original study

design

Protocol Deviations:

- Omissions or deviations (“errors”)

- Should be recorded separately

- Not as retrospective amendments

- Assessed for significance

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SOPs

Histology SOPs:

- Tissue trimming

- Sectioning

- Staining, including special stains

- Equipment maintenance

- Records to be kept

- Slide shipping

- Chain of custody

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SOPs

Guides for organ sampling and trimming in

rats and mice:

http://reni.item.fraunhofer.de/reni/trimming/

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Adrenal Gland Trimming

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- Through cortex

and medulla

- 1 per side

- Longitudinal

Seminal VesicleTrimming

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- Mid section

- 1 per side

- TS, with

coagulating gland

SOPs

Benefits:

- Consistency (between staff, over time)

- Training

- Reconstruction of methods used in past

- Avoids repetitive documentation of

procedures

- Problems only have to be solved once

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Data and Documentation

When the study is completed, the data,

documentation and samples must

completely support what was done

It must be possible for a third party to

reconstruct the study from what remains

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Data and Documentation

Identifies:

- Who did it?

- When was it done?

- What was done?

- What were the results?

- Documents that SOPs and protocol were

followed

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Data and Documentation

Raw Data:

- Original records (or verified copies) of

original observations and activities in the

study

- Recorded directly, promptly, accurately,

legibly and identified by who did it.

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Data and Documentation

Raw Data:

- Original records (or verified copies) of

original observations and activities in the

study

- Recorded directly, promptly, accurately,

legibly and identified by who did it.

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Histology Documentation

- Tissues taken

- Tissues trimmed

- Tissues in block

- Tissues on slide

- Staining procedures

- QC check done

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Histology Quality

Pathologist’s niggles:

- Same blocking pattern every time, e.g.

ovaries/uterine horns/cervix/vagina

- No unnecessary extra sections

- No mixing slides of different thickness

- Same stain colour for control and treated

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Histology Materials

Specimens (wet tissue, blocks, slides)

- Uniquely identified

- Retained

- Retrievable for future queries

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Histology Quality Control

- Check that activities carried out properly

- Check all tissues taken at necropsy

- Check slide identification, sectioning,

staining

- Check tissue on slide

- Check data complete, signed

- Second person does it! Not QA!

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Pathology Raw Data

- Record of the pathologist’s findings for

individual animals (when pathology data

finalised)

- Pathologist’s text interpreting the

findings

- (Possibly) computer file containing

study data

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Pathology Quality Control

Peer Review:

- Review by a second pathologist of draft

report, 10% of animals, all neoplasms,

all target organs.

- Discussion of Peer Review findings

using double header microscope

- Final Report with consensus opinion

between pathologists

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Pathology Quality Control

Peer Review:

- Typically takes two weeks for a

carcinogenicity study

- Data retained: records of animals and

organs examined, consensus achieved

- Not about “correcting” minor variations

between pathologists

- Best result as a collaborative exercise

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Pathology Quality Control

Pathology Working Group (PWG):

- After report finalised, often mouse and

rat studies together

- Study pathologist, reviewing

pathologist, PWG leader, six “Experts”

- Characterise important or controversial

findings and assess their significance to

humans

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Study Report

- Complete and accurate reflection of

protocol and raw data

- One study, one report

- Reviewed and approved by QA

- Signed by responsible people

- Date of report and finalisation is date

signed by study director

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Archive

- All data and specimens retained after

study completion

- Secure, limited access

- Protected against deterioration

- Easily retrievable

- Regulations define period of retention,

but usually held for useful duration of

the compound

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Computers

- Same principles of GLP apply

- Suitable for purpose

- Raw data defined

- Formal validation process of hardware

and software before operational use

- Maintenance, backup, disaster plan,

obsolescence and migration to new

system need to be defined

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Multi-site Studies

Multi-site study

- Where phases are conducted on more

than one site

- Phase: a defined set of activities

- Test Facility: location of Study Director

- Test Sites: locations of other phases

- Principal Investigators: carry out duties

of study director at test sites

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Multi-site Studies

Protocol

- Identify test sites, PI, and phases

- How test site data will be included in

report

SOPs

- Selection and monitoring of test site

- QA procedures

- Chain of custody for data and specimens

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Multi-site Studies

Report

- One report for study

- Include PI contributions

- Include evidence of QA involvement

- Define archive location

- Indicate degree of GLP compliance

- Compliance statement for whole study

signed by Study Director

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Multi-site Studies

Practical issues:

- Communication (Languages!)

- Defining responsibilities

- Assessment of test sites

- Monitoring and reporting of findings

- Chain of custody for samples and

specimens

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Summary

GLP is little more than good scientific

practice, allowing:

- Repeatability

- Reconstruction

- Peer Review

It requires application of the same principles

as any good research:

- Discipline and Care

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