. 2021 Jun 16;32(11-12):563–580. doi: 10.1089/hum.2020.269

Table 2.

Design of the safety studies in nonhuman primates

Parameter	Time of Assessment (Study Week)^a
Parameter	Pre	0	1	2	4	8	13	26	52
Vector/vehicle administration^b		■
Physical examination	■		■		■		■		■
Clinical observations	■	■	■	■	■	■	■	■	■
Weight, rectal temperature, vital signs^c	■	■	■	■	■	■	■	■	■
Complete blood counts^d	■	■	■	■	■	■	■	■	■
Serum chemistry^e	■	■	■	■	■	■	■	■	■
CSF^f		■	■				■	■	■
Anti-AAVrh.10 immunity^g	■	■	■	■	■	■	■	■	■
Behavior^h	■	■	■	■	■	■	■	■	■
MRIⁱ	■						■	■	■
Sacrifice^j			■				■	■	■
Gross and histopathology^k			■				■	■	■

^{^a}

n = 24 (13F/11M). Study day 0 was day of surgery; all observations and safety assessments on day 0 were performed before surgery. Times for assessment for remaining time points were as follows: up to 2 weeks acceptable timeframe was ±1 day and for times over 2 weeks it was ±3 days.

^{^b}

Vector dosing: PBS (n = 2F/2M); AAVrh.10Null, 1.5 × 10¹² gc (n = 3F/1M); AAVrh.10hARSA, 2.85 × 10¹⁰ gc (n = 4F/4M); AAVrh.10hARSA, 1.5 × 10¹² gc (n = 4F/4M). A subset of each group was euthanized at 1, 13, 26, and 52 weeks as detailed in Table 1.

^{^c}

Assessment of general safety parameters, including temperature, pulse, respiratory rate, and weight.

^{^d}

Complete blood tests included: white cell count, red blood cell count, reticulocytes (% and absolute), hemoglobin, hematocrit, mean corpuscular volume, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration, segmented neutrophils (% and absolute), lymphocytes (% and absolute), monocytes (% and absolute), eosinophils (% and absolute), basophils (% and absolute), and platelet count.

^{^e}

Serum chemistry tests included: alkaline phosphatase, alanine aminotransferase, aspartate aminotransferase, gamma glutamyl transferase, albumin, total protein, globulin, total bilirubin, BUN, creatine kinase, triglycerides, glucose, lactate dehydrogenase, calcium, phosphorus, bicarbonate, amylase, lipase, sodium, chloride, potassium, sodium/potassium, albumin/globulin, and BUN/creatinine ratios, anion gap.

^{^f}

CSF was collected at two time points per NHP, before vector administration and at necropsy, for analysis (quantification of ARSA protein and anti-AAVrh.10 total and neutralizing antibodies).

^{^g}

Assessments included: humoral anti-AAVrh.10 capsid immunity by serum total anti-AAVrh.10 and neutralizing AAVrh.10 antibody titers and by CSF total anti-AAVrh.10 titers and CSF neutralizing anti-AAVrh.10 antibody titers.

^{^h}

At the indicated times, behavioral assessments were performed with videotaping at rest and in responses to a series of standardized challenges, with extraction of quantitative traits as previously described.

^ⁱ

MRI was performed on groups described in Table 1.

^{^j}

A subset for each time point as described in Table 1.

^{^k}

Before necropsy, blood and CSF were sampled for analyses as detailed in footnotes d to g. During the necropsy, the animals were observed for pathology, with organs weighed and examined for gross pathology and abnormalities. The brain was excised, divided into hemispheres, and fixed with 10% formalin for histopathology examination of the administration sites. Samples of other organs (liver, lung, biceps, kidney, gonads, seminal vesicles, testes, epididymides, skin, spleen, ileum, heart, multiple sections of the spinal cord, lymph nodes, eyes with lacrimal glands, optic nerve, glossopharyngeal nerve, facial nerve, parotid and submandibular salivary glands, and any gross abnormalities) were collected at necropsy, fixed with formalin for histopathological analysis.

■ = test required for the research study.

BUN, blood urea nitrogen; CSF, cerebrospinal fluid; F, female; M, male.