7 TIPS FOR NAVIGATING THE COMPLEXITIES OF IN VITRO NAMS STUDIES

As a board-certified toxicologist and experienced Good Laboratory Practices (GLP) Consultant, I spend quite a bit of time advising on quality issues of New Approach Methodologies (NAMs) in support of toxicology programs. While these alternative methods are exciting and full of promise, they come with their own set of challenges. 

NAMs have been in development for several decades, and are now an accepted part of the toxicological method landscape. They are reshaping the field by providing proven and in some cases cost-effective ways of testing specific toxicological characteristics of potential products to supplement animal testing. 

But as with any new technology, there are some growing pains. In this blog, I’ll dive into some of the key challenges that you may face when incorporating NAMs into your studies and discuss potential solutions to each one. Grab your lab coat and let’s get to it!

1. DATA INTEGRITY: ENSURING YOUR RESULTS ARE AS SOLID AS YOUR REPUTATION

Obviously, data integrity is the cornerstone of any toxicology study. Whether you're using traditional animal models or cutting-edge in vitro methods, data quality is non-negotiable. And when it comes to NAMs, data integrity can become a challenging issue.

The Issue: In vitro NAMs often rely on complex experimental systems with multiple endpoints that result in the generation of large amounts of data. This can include anything from gene expression changes in cultured cells to high-throughput, molecular interaction screenings. The more data, the more potential for errors, especially if the data is not adequately captured, stored, and analyzed.

The Solution? Partner with a CRO that has adopted strong data management practices that include:

• Electronic laboratory notebooks (ELNs): Using ELNs that are 21 CFR Part 11-compliant ensures that data is securely recorded, tamper-proof, and easily auditable. These systems allow for tracking of everything from raw data to final analysis with time stamps, version control, and user authentication.
• Automated data collection: Validated, automated systems help minimize human error and ensure that your results are captured consistently and accurately. This is crucial when dealing with high volumes of data. It is crucial to make certain that these comply with 21 CFR Part 11.
• Clear standard operating procedures (SOPs): A CRO with SOPs for data management can go a long way in ensuring that everyone on the research and development team is handling data consistently to ensure integrity.
• Reliable quality control (QC) systems: Reliable QC, preferably with daily checks, helps detect discrepancies while they are still fresh.

When your data is properly locked down, you can rest easier knowing that your results will stand up to scrutiny.

2. TRAINING AND PERSONNEL QUALIFICATION: THE RIGHT PEOPLE MAKE ALL THE DIFFERENCE

One benefit of NAMs is that they often require specialized knowledge; whether that’s understanding complex in vitro assays or using advanced software to analyze data. Sponsors need assurance that their CRO partner has fully validated models as well as the expertise and qualified personnel to handle these sophisticated techniques effectively.

The Issue: In vitro methods often require expertise in cell culture, molecular biology, and sometimes even bioinformatics. Without the right personnel, your study could end up as a train wreck rather than the gold standard.

The Solution? Partner with someone that invests in training their team. A strong CRO will have a fully GLP-trained team, with regular workshops and courses to keep teams sharp, especially when working with new technologies. Personnel should be cross-trained in both technical procedures and GLP requirements, equipped to handle unexpected situations with ease and ensuring study continuity for sponsors.

3. EQUIPMENT AND SOFTWARE VALIDATION: A BROKEN MACHINE EQUALS A BROKEN STUDY

Researchers can have the best in vitro assay in the world, but if the equipment or software isn’t qualified or validated, you’re in trouble. Imagine tests being run on an uncalibrated spectrophotometer or feeding your data into a glitchy analysis program; that’s a nightmare no one wants to experience.

Just as importantly, the NAMs models themselves must also be validated to ensure they are scientifically sound for their intended purpose. Without proper model validation, even perfectly functioning equipment can produce misleading or non-translatable results.

The Issue: In vitro NAMs often rely on expensive and sensitive equipment, like high-content screening systems, flow cytometers, or automated liquid handlers. If these systems are not validated according to GLP standards, data integrity could be compromised.

The Solution?

• Routine calibration and validation: Equipment should be calibrated regularly, and software should undergo validation to ensure it performs as expected under different conditions. Ensure your CRO maintains up-to-date calibration records to avoid issues during inspections.
• Computerized systems validation: Any software used to analyze or store data must be validated according to 21 CFR Part 11. This includes, but is not limited to, installation qualification, operational qualification, and performance qualification. 
• Model validation: In vitro NAMs models should undergo rigorous scientific validation to demonstrate that they are predictive, reproducible, and appropriate for the intended regulatory or research application. This includes biological relevance, sensitivity, specificity, and consistency across different test runs. 

Without the right equipment and software in top shape, you run the risk of compromising your study data.

4. STUDY RECONSTRUCTION: WHEN THINGS GO OFF THE RAILS

One of the realities of toxicology studies is that things sometimes go wrong. It could be anything from human error to a batch of contaminated cell lines. When that happens it’s essential to be able to reconstruct the study; and that’s where traceability and documentation come into play.

The Issue: If something goes wrong in the middle of a study, is the CRO able to consult the original data and figure out what happened? Poor documentation or inadequate data management can make this process nearly impossible.

The Solution?

• A CRO that keeps diligent record keeping: This includes raw data, notes on protocol deviations, and observations during the experiment. If a Part 11-compliant EDC system is used, it should easily allow tracing of any changes, modifications, or discrepancies.
• Audit trails: Data systems should include audit trails that track every action taken, from the moment data is entered, to the final analysis. This ensures that you and the CRO can identify where things went wrong, if necessary.

Study reconstruction is one of those things you hope you never need, but it’s best to have a CRO partner that is always prepared. A little foresight can save a lot of headaches later.

5. ARCHIVING: BUILDING A LASTING PAPER TRAIL

Now, let’s talk about one of the least exciting, but most crucial aspects of good laboratory practices: archiving. If you think your data can be stored on a few USB drives in a drawer, think again. Regulatory authorities expect secure, long-term storage of all study records.

The Issue: Storing data properly is a challenge, especially when dealing with massive datasets from in vitro studies. Losing or improperly archiving data is not just a minor error—it could put the whole study at risk.

The Solution? 

• Long-term digital storage: You CRO should use secure, compliant digital storage solutions that can handle the volume of data and keep it accessible for years. This might involve cloud storage with robust backup systems, or local servers with redundancy.
• Physical archiving: For any physical records, your CRO should ensure they are stored in a secure, organized manner, allowing for easy retrieval.

Secure data storage is always worth the effort.

6. SPONSOR EXPECTATIONS AND COMMUNICATIONS: MANAGING THE CLIENT-CONSULTANT-CRO RELATIONSHIP

As a sponsor with toxicology studies, it's natural to have high expectations for results. However, it's important to recognize that there can be challenges and complexities associated with the use of NAMs. 

The Issue: Sponsors may be eager to adopt NAMs but might not fully recognize the complexity and expense required to produce reliable results. By being aware of these realities, we can work together to navigate any potential obstacles and achieve meaningful outcomes.

The Solution?

• Set clear expectations: Your CRO must be upfront about timelines, costs, and potential challenges. Using NAMs may take time to get right, and regulatory acceptance isn’t guaranteed (yet).
• Justify differences: Have your CRO determine that the differences in cost, time, and data requirements associated with using NAMs, instead of traditional methods, are justified and aligned with your goals and objectives.
• Frequent communication: Your CRO should keep your and all stakeholders in the loop with regular updates on progress, challenges, and milestones. Transparency will help you avoid surprises at the end of the study. 

Good communication builds trust, and trust makes everything run smoother.

7. REGULATORY ACCEPTANCE UNCERTAINTY: THE WILD CARD

The final hurdle in NAMs is regulatory acceptance. Regulatory bodies like the FDA, EPA, and OECD-member countries are gradually adopting NAMs, but a lot of uncertainty remains about which methods will be accepted in various jurisdictions.

The Issue: We are still far from having validated NAMs that can fully replace traditional toxicology studies, due to the extensive validation required, and the large number of models that need to be assessed and approved. Will your study’s data be accepted as a supplement to traditional animal studies? Are you sure the method is robust enough for regulatory approval?

The Solution?

• Stay informed: Keep up with the latest developments in regulatory guidance. The OECD, for example, provides regular updates on validated methods and acceptance criteria. The FDA also issues numerous guidelines. 
• Engage with regulators: Early engagement with regulatory agencies can help you understand their expectations and provide input on the acceptability of the methods. This can help clear up any confusion and set your program on the right track.
• Consider Whole-Animal Complexity: Always appreciate that in nature, the whole animal (including humans) is always more than the sum of the parts. NAMs focus on specific aspects of toxicity and cannot predict overall systemic conclusions comprehensive to the whole animal.
• Remain patient: While there’s still uncertainty, things are moving in the right direction. Patience and persistence are key.

A CRO experienced in NAMs can help interpret evolving guidelines and design studies that balance innovative approaches with regulatory expectations, giving you the confidence that your study results will be recognized and trusted.

THE BOTTOM LINE

For sponsors, adopting NAMs in toxicology programs can feel like navigating a maze for, but partnering with experienced CROs that pay careful attention to data integrity, you’ll be well on the right path. Stay proactive, set clear expectations with your CRO partners, and follow guidance as it evolves—NAMs are here to stay and will make toxicology testing smarter, safer, and more efficient while helping to supplement animal studies.

ABOUT THE AUTHOR

Robin Guy is a Board-Certified Toxicologist (DABT) and a Registered Quality Assurance Professional in Good Laboratory Practices (RQAP-GLP). With 19 years of

prior industry experience, she has spent an additional 25 years consulting, helping clients navigate the intricate world of GLP compliance and toxicology. Her gardening skills, however, have been honed for far longer, and it is rumored she can identify a toxic plant with just one glance.

Robin's expertise spans a wide range of services, including administering GLP training courses, conducting audits, and helping labs transform into compliant, inspection-ready facilities. Robin also assists clients in addressing regulatory citations and providing ongoing inspection training and follow-up. When it comes to toxicology, Robin is deeply involved in the design, monitoring, and reporting of GLP toxicology studies, ensuring that nonclinical summaries are crafted for everything from GRAS to IND to NDA submissions.

A long-time contributor to the Society of Quality Assurance, Robin has served on the Program Committee for 15 years and twice chaired the Quality Assurance Consulting Specialty Section. She is also a proud Past President of the American College of Toxicology (ACT).

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_{This blog was first published in October 20205.}