Federal Register / Vol. 50, No. 160 / Monday, August 19, 1985 / Notices 
33465 
vector, and the delivery vector (if any). 
Provide complete sequence analysis or a 
detailed restriction map of the total 
construct. 
(2) What regulatory elements does the 
construct contain (e.g., promoters, 
enhancers, polyadenylation sites, 
replication origins, etc.)? 
(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 (nucleic acids, 
proteins, etc.) or contaminating viruses 
or other organisms in the cells or serum? 
(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 
laboratory animals) for claims about the 
efficacy and safety of the proposed 
system for gene delivery. 
a. Laboratory studies of the delivery 
\ system. (1) What cells are the intended 
i recipients of gene therapy? If recipient 
cells are to be treated in vitro 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 treated cells will act as recipients? 
(2) Is the delivery system efficient? 
What percentage of the target cells 
contain the inserted DNA? 
(3) How is the structure of the added 
DNA sequences monitored and what is 
the sensitivity of the analysis? Is the 
added DNA extrachromosomal or 
integrated? Is the added DNA 
unrearranged? 
(4) How many copies are inserted per 
cell? How stable is the inserted DNA 
both in terms of its continued presence 
and its structural stability? 
b. Laboratory studies of gene 
expression. Is the inserted gene 
expressed? To what extent is expression 
only from the desired gene (and not from 
the surrounding DNA)? In what 
percentage of cells does expression 
occur? Is the product biologically active? 
What percentage of normal activity 
results from the inserted gene? Is the 
gene expressed in cells other than the 
target cells? If so, to what extent? 
c. Laboratory studies pertaining to the 
safety of the delivery /expression 
system. (1) If a retroviral system is used: 
(a) What cell types have been infected 
with the retroviral vector preparation? 
Which cells, if any, produce infectious 
particles? 
(b) How stable are the retroviral 
vector and the resulting provirus against 
loss, rearrangement, recombination, or 
mutation? What information is available 
on how much rearrangement or 
recombination with endogenous or other 
viral sequences is likely to occur in the 
patient's cells? What steps have been 
taken in designing the vector to 
minimize instability or variation? What 
laboratory studies have been performed 
to check for stability, and what is the 
sensitivity of the analyses? 
(c) What laboratory evidence is 
available concerning potential harmful 
effects of the treatment, e.g., 
development of neoplasia, harmful 
mutations, regeneration of infectious 
particles, or immune responses? What 
steps have been taken in designing the 
vector to minimize pathogenicity? What 
laboratory studies have been performed 
to check for pathogenicity, and what is 
the sensitivity of the analyses? 
(d) Is there evidence from animal 
studies that vector DNA has entered 
untreated cells, particularly germ line 
cells? What is the sensitivity of the 
analyses? 
(e) Has a protocol similar to the one 
proposed for a clinical trial been carried 
out in non-human primates and/or other 
animals? What were the results? 
Specifically, is there any evidence that 
the retroviral vector has recombined 
with any endogenous or other viral 
sequences in the animals? 
(2) If a non-retroviral delivery system 
is used: What animal studies have been 
done to determine if there are 
pathological or other undesirable 
consequences of the protocol (including 
insertion of DNA into cells other than 
those treated, particularly germ line 
cells)? How long have the animals been 
studied after treatment? What tests have 
been used and what is their sensitivity? 
3. Clinical Procedures, Including Patient 
Monitoring 
Describe the treatment that will be 
administered to patients and the 
diagnostic methods that will be used to 
monitor the success or failure of the 
treatment. If previous clinical studies 
using similar methods have been 
performed by yourself or others, 
indicate their relevance to the proposed 
study. 
a. Will cells (e.g., bone marrow cells) 
be removed from patients and treated in 
vitro in preparation for gene therapy? If 
so, what kinds of cells will be removed 
from the patients, how many, how often, 
and at what intervals? 
b. Will patients be treated to 
eliminate or reduce the number of cells 
containing malfunctioning genes (e.g.. 
through radiation or chemotherapy) 
prior to gene therapy? 
c. What treated cells (or vector/DNA 
combination) will be given to patients in 
the attempt to administer gene therapy? 
How will the treated cells be 
administered? What volume of cells will 
be used? Will there be single or multiple 
treatments? If so, over what period of 
time? 
d. What are the clinical endpoints of 
the study? How will patients be 
monitored to assess specific effects of 
the treatment on the disease? What is 
the sensitivity of the analyses? How 
frequently will follow-up studies be 
done? How long will patient follow-up 
continue? 
e. What are the major potential 
beneficial and adverse effects of 
treatment that you anticipate? What 
measures will be taken in an attempt to 
control or reverse these adverse effects 
if they occur? Compare the probability 
and magnitude of potential adverse 
effects on patients with the probability 
and magnitude of deleterious 
consequences from the disease if gene 
therapy is not performed. 
f. If a treated patient dies, what 
special studies will be performed as part 
of the autopsy? 
4. Public-Health considerations 
Describe any potential benefits and 
hazards of the proposed therapy to 
persons other than the patients being 
treated. Specifically: 
a. On what basis are potential public 
health benefits or hazards postulated? 
b. Is there a significant likelihood that 
the inserted DNA will spread from the 
patient to other persons or to the 
environment? 
c. Will any precautions be taken 
against such spread (e.g., to patients 
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