Federal Register / Vol. 55, No. 41 / Thursday, March 1, 1990 / Notices 
appropriateness of gene therapy of somatic 
cells in humans for specific genetic diseases. 
Somatic cell gene therapy is seen as an 
extension ofrpresent methods of therapy that 
might be preferable to other technologies. 
' In light of this public support, the RAC is 
prepared to consider proposals for somatic 
cell gene therapy. 
(9) In their evaluation of proposals 
involving the transfer of recombinant DNA 
into human subjects, the RAC and its 
Subcommittee will consider whether the 
design of such experiments offers adequate 
assurance that their consequences will not go 
beyond their purpose, which is the same as 
the traditional purpose of clinical 
investigations, namely, to protect the health 
and well-being of the individual subjects 
being treated while at the same time 
gathering generalizable knowledge. 
Two possible undesirable consequences of 
the transfer of recombinant DNA would be 
unintentionah (1) Vertical transmission of 
genetic changes from an individual to his or 
her offspring or (2) horizontal transmission of 
jviral infection to other persons with whom 
the individual comes in contact Accordingly, 
this document requests information that will 
enable the RAC and its Subcommittee to 
assess the possibility' that the proposed 
experiments will inadvertently affect 
reproductive cells or lead to infection of other 
people (e.g., treatment persoimel or relatives]. 
(10) In recognition of the social concern 
that surrounds the subject of gene transfer, 
the Subcommittee will cooperate with other 
groups in assessing the possible long-term 
consequences of the tra^fer of recombinant - 
DNA into human subjects and related 
laboratory and animal experiments in order 
to define appropriate human applications of 
this emerging technology. 
(11) Responses to the questions raised in 
these Points to Consider should be provided . 
in the form of either written answers or 
references to spedhc sections of the protocol 
or its appendices. . 
'>](12] investigators should indicate points 
wUch are not applicable with a brief 
explanation. Investigators submitting 
proposals that employ essentially the same 
vector systems (or with minor variations], 
and/or that are based on the same preclinical 
testing as proposals previously reviewed by 
the Subcommittee and the Recombinant DNA 
Advisory Committee (RAC), may refer to 
precedirig documents without having to 
rewrite such material 
L Description of Proposal 
A. Objectives and rationale of the 
proposed research. 
State concisely the overall objectives and 
rationale of the proposed study. Please 
provide information on the specihc points 
that relate to whichever type of research is 
being proposed: 
l.Use of recombinant DNA for therapeutic 
purposes 
^r.Porjesearch in which recombinant DNA is 
transferred in order to treat a disease or 
disorder (e.g., genetic diseases, cancer, and 
metabolic diseases], the following questions 
.should be addressed: 
a. Why is the disease selected for 
treatment by means of gene therapy a good 
candidate for such treatment? 
b. Describe the natural history and range of 
expression of the disease selected for . 
treatment What objective and/or 
quantitative measures of disease activity are 
available? In your view, are the usual effects 
of the disease predictable enough to allow for 
- meaningful assessment of the results of gene 
therapy? 
a Is the protocol designed to prevent all 
manifestations of the disease, to halt the 
progression of the disease after symptoms 
have begun to appear, or to reverse 
manifestations of the disease in seriously ill 
victims? 
d. What alternative therapies exist? In 
what groups of patients are these therapies 
effective? What are their relative advantages 
and disadvantages as compared with the 
proposed gene therapy? 
2. Transfer of DNA for Other Purposes 
a. Into what ceils will the recombinant 
DNA be transferred? Why is the transfer of 
recombinant DNA necessary for the proposed 
.research? What questions can be answered 
by using recombinant DNA? - 
b. What alternative methodologies exist? 
What are their relative advantages and 
disadvantages as compared to the use of 
recombinant D.NA? 
B. Research design, anticipated risks and 
benefits. 
1. Structure and characteristics of the 
biological system 
Provide a full description of the methods 
and reagents to be employed for gene 
delivery and the rationale for their use. The 
foUowing are spedffc points to be addressed: 
A. What is the structure of the cloned DNA 
that will be used? 
(1] Describe the gene (genomic or cDNA], 
4he bacterial plasmid or phage vector, and the 
delivery vector (if any]. Provide complete 
nucleotide sequence analysis or a detailed 
restriction enzyme map of the total construct 
(2] What re^atory elements does the 
construct contain (e.g., promoters, enhancers, 
polyadenylation sites, replication origins, 
etc.]? From what source are these elements 
derived? Summarize what is currently known 
about the regulatory character of each 
element 
(3] Describe the steps used to derive the 
DNA construct 
b. What is the structure of the material that 
will be administered to the patient? 
. (1] Describe the preparation, structure, and 
composition of the materials that will be 
given to the patient or used to treat the 
patient's cells. 
(a] If DNA, what is the purity (both in 
terms of being a single DNA species and in 
terms of other contaminants]? What tests 
•have been used and what is the sensitivity of 
the tests? 
(b] If a virus, how is it prepared from the 
DNA construct? In what cell is the virus 
grown (any special features]? What medium 
and serum are used? How is the virus 
purified? What is its structure and purity? 
What steps are being taken (and assays used 
with their sensitivity] to detect and eliminate 
any contaminating materials (for example, - 
7-143 
VL30 RNA. other nucleic acids, or proteins] 
or contaminating viruses (both replication- ' 
competent or replication-defective] or other 
organisms in the cells or serum used for 
.preparation of the v^s stock including any 
contaminants that may have biological 
effects? 
(c] If co-cultivation is employed, what 
kinds of cells are being used for co- 
cultivation? W'hat steps are being taken (and 
assays used with their sensitivity] to detect 
and eliminate any contaminating materials? 
Specifically, what tests are being done to 
assess the material to be returned to the 
patient for the presence of live or killed doner 
cells or other non-vector materials (for 
example. VL30 sequences] originating from 
those cells? 
(d] If methods other than those covered by 
(a)-(c] are used to introduce new genetic 
information into target cells, what steps are 
being taken to detect and eliminate any 
contaminating materials? What are possible 
sources of contamination? What is the 
sensitivity of tests used to monitor 
contamination? 
(2) Describe any other material to be used 
in preparation of the material to be 
administered to the patient. For example, if a 
viral vector is proposed, what is the nature of 
the helper virus or cell line? If carrier 
particles are to be used, what is the nature of 
these? 
2. Preclinical studies, including risk- 
assessment studies 
“Describe the experimental basis (derived 
from tests in cultured cells and animals] for 
claims about the efficacy and safety of the 
proposed system for gene delivery and 
explain why the model(s] chosen is (are] the 
most appropriate. 
a. Laboratory studies of the delivery 
system. 
■. (1] What cells are the intended target cells 
of recombinant DNA? It target cells are to be 
treated ex vivo and returned to the patient, 
how will the cells be characterized before 
. and after treatment? What is the theoretical 
and practical basis for assuming that only the 
target ceUs will incorporate the DNA? 
(2] Is the delivery system efficient? What 
percentage of the target cells contain the 
added DNA? 
(3] How is the structure of the added DNA 
sequences monitored and what is the 
sensitirity of the analysis? Is the added DNA 
extra chromosomal or integrated? Is the 
added DNA urmearranged? 
(4] How many copies are present per cell? 
How stable is the added DNA both in terms 
of its continued presence and its structural 
stability? 
b. Laboratory studies of gene transfer and 
expression. 
(1] What animal and cultured cell models 
were used in laboratory studies to assess the 
in vivo and in vitro efficacy of the gene 
' transfer system? In what ways are these 
models similar to and different from the 
proposed human treatment? 
(2) What is the minimal level of gene 
transfer and/or expression that is estimated 
to be necessary for the gene transfer protocol 
to be successful in humans? How was this 
level determined? 
Recombinant DNA Research, Volume 14 
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