NIH All About Grants Podcast on Considering Alternatives to Animals in Applications

>>>� From the National Institutes of Health in Bethesda, Maryland, this is "All about Grants."�

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>> Kosub:� Hello and welcome to another edition of NIH's "All about Grants" podcast.� I'm your host, David Kosub, with the NIH's Office of Extramural Research, and today we'll be discussing a topic that's important when you're putting together your application if you're proposing the use of animals, and that's also considering the use of alternatives to animals in your research.�

And with us we have Dr. Neera Gopee.� She directs the Policy and Education Office within the NIH's Office of Laboratory Animal Welfare, and we also have Dr. Christine Livingston.� She is a scientific review officer with the National Center for Advancing Translational Sciences, or NCATS. Thank you both for being with us.�

>> Gopee:� Thank you, David.� It's good to be here.�

>> Livingston:� Thanks for having us.� Glad to be here.

>> Kosub:� Great, great. So, Neera, I'm going to turn to you first.� I want you to help frame the conversation, specifically telling us around, like, the three Rs ‑‑ replacement, refinement, and reduction ‑‑ in the use of laboratory animals and also how NIH considers these alternatives in research.

>> Gopee:� So, David, this term ‑‑ this concept of three Rs was coined way back in 1949 by Drs. Russell and Burch, where they advocated new scientific approaches by introducing the concept of the three Rs ‑‑ reduction, refinement, and replacement.� And the three Rs are used as an ethical framework for improving laboratory animal welfare and is synonymous with alternatives or alternative methods for minimizing potential for pain and distress in animals used in biomedical research.

>> Kosub:� All right.� Thank you for that. So are there any regulations or policies that must be taken into consideration for the use of alternatives to animals?�

>> Gopee:� Actually, there are numerous federal laws and regulations and policies that require that research involving the use of animals must be reviewed and determined to be scientifically and ethically justifiable, with considerations of alternatives to animal research.� And the NIH actually conducts research using animals at 23 institutes and funds extramural research that involves the use of animals.� I won't go into each one of these policies and regulations, but as this podcast progresses, we may mention a few of these.

> Kosub:� So along those lines, how can we ensure that researchers do consider the use of alternatives to animals?�

>> Gopee:� So I would say that first of all, the Health Research Extension Act of 1985, which provides legislative mandate for the PHS policy ‑‑ it requires researchers involved with animal care, treatment, and use ‑‑ that they have available to them instruction or training in the concept of availability and use of research or test methods that limit the use of animals or limit animal distress.�

In addition, the PHS policy also requires institutions to describe in their assurance agreement with NIH OLAW a synopsis of training or instruction offered to scientists, animal technicians, and other personnel involved in the care and use of animals.� This includes the humane practice of animal care and use and also to describe methods that minimize the number of animals required to obtain valid results and minimize animal distress.

I'd also like to mention that the U.S. Government Principle Number Three, which is a group of nine ethical principles governing the care and use of animals used in research, requires researchers to use the minimum number of animals required to obtain valid results and to consider alternative methods such as mathematical models, computer simulation, and in vitro biological systems.� And the principles call for researchers to consider alternatives, regardless of whether the procedures cause pain or distress to animals.�

And finally, I'd like to also mention that as a condition of receiving funding for animal research, institutions and researchers must comply with the Eighth Edition of the Guide for the Care and Use of Laboratory Animals, also known as "The Guide."� And The Guide states that in preparing and reviewing research protocols, researchers in animal care and use committees should consider the availability or appropriateness of using less invasive procedure, other species, isolated organ preparation, cell or tissue culture, or computer simulation, and The Guide does not limit this research to painful or distressful procedures.�

I'd like to also ‑‑ please note that researchers must comply with all other federal statutes and regulations relating to animals.

>> Kosub:� And what about for the institutional side?�

>> Gopee:� So great question, David. From an institutional perspective, no ‑‑ absolutely no animal work can be conducted without an approved animal welfare assurance, which is pretty much, in essence, a contract between NIH and the institution which commits them to good animal welfare.� So that's something that must be done by the institution, by the researcher, and must be negotiated between OLAW and that institute ‑‑ the granted institution.� And within that animal welfare assurance, there is a description of the training or the instruction available to scientists, animal technicians, and other personnel who are involved in animal care, which includes training that minimizes the number of vertebrate animals used, as well as the minimization of pain and distress.

I'd also like to add that NIH OLAW also provides training to researchers in animal care and alternatives, and we do have ‑‑ under our "useful links" tab on OLAW's web page, there is searches for alternatives as well as database searches.� OLAW also does site visits.� We conduct site visits to assess the adequacy and accuracy of the institution's compliance with the PHS policy.� And so by doing that, when ‑‑ during our site visit, what is included in that site visit is a review of the IACUC�s approved protocol form, which includes a search for alternatives.� So there's a whole range of different checks and balances in place to ensure that the institutions do consider the use of alternatives, as well as NIH's oversight of these alternative searches.

>> Kosub:� Wonderful.� Great.� So now I actually want to turn to Christine. Can you tell us where in the application someone might be talking about alternatives to animals and how they might justify the use of animals if there are no alternatives?�

>> Livingston:� So that would be included with the rationale for your research.� It would be located in the research plan segment of your application.� That's the 6‑ or 12‑page research plan that's associated with any grant application coming into the NIH.� In that section of the application, you would ‑‑ if there are alternatives, you would probably be using them.� If there are no alternatives, then you need to justify your choice of animal model and not only your choice of animal model but if there are models that are close to the one that you are using.� You might cite those but indicate what their limitations are.� Basically you're providing the rationale for your scientific approach, and that includes your animal or non‑animal model.� It should all be up in the research plan.

You would probably also justify the use of the number of animals that you would be using in the research plan, so you would probably indicate somewhere in the rigor and reproducibility section the number of animals you're using.� And you will minimize the number of animals you're using but not to the point that you would not get a valid scientific answer.� So you want to use enough animals to get a good answer but not more than you need.�

So your choice of animal model and the number of animals would all be justified under "rationale" in the research plan.

>> Kosub:� Thank you for that. So for those animals that are used, how can a researcher indicate the methods that are being used ‑‑ actually do consider‑‑ have considerations for pain and any sort of distress on those that are involved?

>> Gopee:� So I can chime in here. To add in to what Christine mentioned, the NIH also requires information on the involvement of vertebrate animals to be submitted by extramural researchers as part of their peer‑review process, what is called a "Vertebrate Animal Section," or VAS.� And this VAS ‑‑ applicants must be complete three criteria within this VAS.� One of these criteria, in addition to requiring that the researchers address alternatives ‑‑ the justification for alternatives and use of alternative models, they must also address and complete a description of the plan interventions to minimize discomfort, distress, pain, and injury, including the use of analgesia or anesthesia, sedation, and ‑‑ as well as a description of humane endpoints.�

>> Kosub:� Great.� Thanks for that. Christine, turning back to you. You're at NCATS.� Neera touched on some of the alternatives earlier, but I wanted to ‑‑ I specifically have a question on that, and what are some alternatives that researchers can consider?�

>> Livingston:� So I work at an institute that advances translational studies, and so we're very interested in that ‑‑ sort of the going from bench science into clinical science, the translation between those.� So we're very interested in drug development and therapeutic development and what are the methods that we can use to do that.� Other methods would be, in some cases, the use of human subjects.� There's not a lot that you can do in basic science or pre‑clinical research where you would use human subjects.� There may be a few things.� But the two big areas are in vitro models or in silico models.� And the in vitro models could cover everything from cell culture, and those cell cultures could be based on primary human cells or human cell lines.� It could be animal cell lines, stem cells.� Could be any sort of thing.�

The ‑‑ two of the big models that have become very popular in terms of in vitro models are the organoids and the microphysiological systems.� The microphysiological systems, in particular, were developed as a way to ‑‑ they were really developed in conjunction with the FDA and DARPA in ‑‑ there was much interest from the European scientific community in developing these.� They're often multicellular ‑‑ can't call them "mini organoids," but they're the critical cells from different organs.� You may take the critical cell types from a liver, for instance, and grow them on a microfloritic platform, and often times those can be grown from human cells or human‑induced pluripotent stem cells, or they may be ‑‑ you may do it with ‑‑ you may develop them with bio printing.� But if they include enough of the critical cell types from a given organ, you can actually use that organ, grow it under conditions that it puts them in both the (indiscernible) as well as physical environment.�

So in other words, if you were looking at lung tissue, you would give it rhythmic stretch.� If you're looking at something vascular or you're looking at something related to the kidney, you would develop your model so that you have shear flow.� And these ‑‑ you put the stem cells on these platforms, and they can actually grow up into really pretty well functioning organs.� And they become wonderful models for testing toxicity and efficacy of different therapeutics.� They're already ‑‑ have a lot of attention from regulatory agencies, such as the FDA, agencies within the European Union, and they're a great way to either reduce ‑‑ or ultimately the goal would be to replace the use of animals for many of the tox efficacy studies that we currently conduct.

The last area of alternatives are the in silico models, and of course those are the mathematical and computer models.� Many of those are dependent on having the correct information and data and variables to plug in in the front end of the model.� So all of the above still have limitations, but they are all working towards reducing the number of animals used and ultimately, as we accumulate more and more data and science and information, to replace the use of animals for many studies.

>> Kosub:� Great, great.� Thank you for that. Neera, I want to jump back on something you mentioned in one of your earlier answers, and that's the IACUC, the Institutional Animal Care and Use Committee. Can you tell us more about their role in this process?�

>> Gopee:� Sure.� So the PHS policy requires that researchers submit written description of their projects to their IACUCs, their Institutional Animal Care and Use Committees.� And included in that written description, they must include a description of research activities that did not necessarily duplicate previous experiments.� They must also include a description of methods and sources used to determine that alternatives were not available.� And so once these descriptions are included in those protocols, the IACUC is responsible for reviewing and approving these descriptions as submitted by the researchers.

>> Kosub:� Well, on a similar vein, what about for peer reviewers? Christine, you're in scientific review.� Can you tell us how the peer reviewers are thinking about alternatives?�

>> Livingston:� So the peer reviewers are also looking at the animal care use protocol.� The big difference is that the peer reviewers are often times individuals who are in the same scientific field as the grant applicant, so they are very familiar with the various animal models or non‑animal models ‑‑ the alternatives.� They know the pluses and the minuses, the advantages, the disadvantages, and the limitations of those models, and they're very good at, number one, evaluating ‑‑ because they're in the same field as the grant applicant, they're very good at evaluating the importance of the question being asked.� Is this really a valid question that we need to ask and address?� They know if the animal model is the best model, if that's what's being proposed.� They can evaluate that.� They know if there's an alternative because they're in that same field.

They also have a different perspective as they look at power analysis and looking at the required number of animals.� They can judge a little bit more ‑‑ I guess their judgment is more informed because they're in the same field ‑‑ looking at the realistic ‑‑ how realistic is the power analysis in terms of the numbers that go into it. So they're good at recognizing your rationale for choice of model, whether it's an animal model or an alternative model.� They're very good at looking at that, and they do speak up on that if they feel that you've chosen a model that is not the best choice scientifically.

>> Kosub:� And what if an application makes it all the way through all these steps and actually gets awarded?� How should alternatives to animals be thought of post award?�

>> Livingston:� I would certainly think that if post award ‑‑ you've been awarded ‑‑ say you've been awarded a five‑year RO1 grant and two or three years down the line a new technique or a new approach comes out or a new model comes out, if you can still achieve your specific aims using an alternative, if I were a grantee, I would certainly pick up the phone, call my program officer, and discuss going in a slightly different direction.� As long as you're still achieving your aims, you certainly could use an alternative model.

>> Gopee:� And I would like to add, David, that, you know, the IACUC is also required to conduct a complete review at least once every three years.� So if there has been in any improvements or progress made in terms of a more appropriate animal model, then that's time where researcher can surely ‑‑ can make changes to their protocol to reflect those advancements in the models used.

>> Kosub:� Great. So before we close out, I would like to give the guests an opportunity for any final thoughts you would like to leave with the audience, and I welcome to both of you guys to mention anything about alternatives to animals that our audience should be thinking about.

>> Gopee:� So I would like to say that, in closing ‑‑ that investigators are accountable for the protection of research animals in their care from the early stages of planning all the way through study completion and that the only acceptable research involved in animals is one that uses the smallest number of animals and causes the least possible pain or distress, as well as a research which is consistent with achievement of a justifiable scientific purpose which is necessary because there's no other way of achieving that purpose and that, you know, the use of laboratory animals are critical to biomedical research.� And alternatives should always be considered to reduce the number of animals used but at this point in time cannot completely replace them.

>> Livingston:� I would like to echo that, and I encourage all investigators to keep ‑‑ to kind of keep their eyes open for alternatives.� There's certainly multiple prominent institutions in the United States that have centers that are looking for the use of alternative methods, and there's certainly European agencies.� I think one of the things I mentioned was the use of animals for toxicity and efficacy testing.� We really would like to see the use of animals in those areas reduced.�

There is an interagency coordinating committee on the validation of alternative methods.� It's called "ICCVAM" for short.� That is actually ‑‑ it's a permanent committee of NIEHS.� It was established in 2000 by a federal authorization act.� It includes representatives from 16 U.S. federal agencies, and each of them is interested in regulatory affairs.� And they look carefully at the use of animals and generation or dissemination of toxicological and safety testing information.� I think it's ‑‑ keeping an eye on those sorts ‑‑ the activities of those agencies is a good idea for all investigators, and I think we owe animals respect.� They have taken us very far in research in the last 150 years, and they warrant respect and good treatment.� And they generate good data when taken good care of and handled well.

>> Kosub:� Great.� Thank you very much, both Neera and Christine, for this opportunity to hear more about how NIH considers the use of alternatives to animals in research. I welcome the audience to check out the Office of Laboratory Animal Welfare's web page for more information about alternatives.� They have some very useful links there. And with that, thank you very much.� ������ This has been David Kosub with another virtual edition of NIH's "All about Grants." Thank you.

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