All animal experiments were in compliance with Institutional Animal Care and Use Committee of Baylor College of Medicine. Nude mice [Athymic Nude-Foxn1nu] and SCID/Beige mice [C.B-17/IcrHsd-Prkdc scid Lyst bg-J] were purchased from Envigo. Osx1-GFP-cre/iDTR was generated from Osx1-GFP-cre [B6.Cg-Tg(Sp7-tTA,tetO-EGFP/cre)1Amc/J] and STP-iDTR mice [C57BL/6-Gt(ROSA)26Sortm1(HBEGF) Awai/J] originally obtained from Jackson Laboratory. For all in vivo experiment, 4- to 7-week-old female mice were used.
C b 17 icrhsd prkdc scid lyst bg j
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The C.B-17/IcrHsd-Prkdc scid Lyst bg-J is a laboratory animal model. It is a genetically modified mouse strain.
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Nod cg rag1tm1mom il2rgtm1wjl szj, by Jackson ImmunoResearch (2 mentions)
Hsd athymic nude foxn1nu, by Inotiv (2 mentions)
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Athymic nude foxn1nu, by Inotiv (1 mentions)
C57bl 6 gt rosa 26sortm1 hbegf awai j, by Jackson ImmunoResearch (1 mentions)
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7 protocols using c b 17 icrhsd prkdc scid lyst bg j
1
Xenograft Mouse Models for Cancer
2
Niraparib Treatment of Patient-Derived Xenograft
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All animal studies were performed under protocols approved by the Mayo Clinic Animal Care and Use Committee. Utilizing previously described methods (34 (link)), PDX PH039 was established intraperitoneally in five female SCID Beige mice (C.B-17/IcrHsd-PrkdcscidLystbg–J; Envigo, Indianapolis, IN). When tumor size (largest cross-sectional area) as monitored by transabdominal ultrasound (SonoSite S-Series Ultrasound, Fujifilm SonoSite, Bothell, WA) reached 0.5–0.8 cm2, mice were randomized to the niraparib arm (n = 3) or source tumor arm (n = 2) as indicated in Figure 1A . On the source tumor arm, tumors were monitored for growth twice weekly by ultrasound, harvested when they reached predefined size criteria, and re-established in two mice. Tumor was passaged five times to control for any changes unrelated to niraparib treatment. On the niraparib arm, mice treated with 100 mg/kg niraparib (dissolved in 0.5% methylcellulose) daily for 21 days via oral gavage were monitored for regrowth of tumors, which were harvested and re-established in three mice. Once established, the tumor was again treated with niraparib for 21 days. This cyclic drug exposure continued until the tumor grew through treatment in two sequential passages. Additional details of the animal studies are described in the Supplementary Methods.
3
Orthotopic Thyroid Tumor Model in Mice
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Assessing Tumor Cell Extravasation
5
Xenograft Tumor Growth Monitoring
6
In Vivo Xenograft Tumor Dissociation and iBH3 Analysis
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All mice were maintained within the DCFI animal facility and all experiments involving animals were conducted in accordance with the DFCI policy and animal protocol, reviewed and approved by the DFCI Institutional Animal Care and Use Committee. These mice were housed in vented caging systems in a 12 h light/12 h dark environment and maintained at uniform temperature and humidity. PC9 cell line xenografts were grown by subcutaneous injection of 5 × 106 PC9 cells in 0.2 mL of 1:1 RPMI:Matrigel into the mid-dorsal flank of 8 week old female SCID-beige mice (C.B-17/IcrHsd-PrkdcscidLystbg-J; Envigo). When tumors grew to ~ 600 mm3 mice were randomized into 3 groups of 4 mice per group. The groups were (1) vehicle, (2) 100 mg/kg venetoclax or (3) 100 mg/kg navitoclax. Twenty-four hours after drug were administered, mice were sacrificed and tumors harvested. Tumors were dissociated as previously described. Then iBH3 was carried out as previously reported by Ryan et al. [18 (link)].
7
Quantifying Tumor Cell Extravasation in Lungs
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Extravasation efficiency assay was performed as described previously [17] . GFP expressing C8161.9 cells with pLKO.1-scrambled, TAO3 knockdown (KD) or rescued (SR) were injected intravenously in tail veins of 8 week old SCID/Beige female mice (C.B-17/IcrHsd-Prkdc scid Lyst bg-J, ENVIGO). 24 hours after injection of 2.5 x 105 of cells, mice were euthanized and tumor cells in lung blood vessels were removed by perfusion, then lungs were harvested. Frozen serial sections of all lungs (50 m thickness) were taken every five sections. For the efficiency analysis, slides were counter stained with Hoechst for 15 min before mounting. All sections were scanned by Zeiss AxiaScan system with ZEN Blue software. GFP positive cells were counted from all scanned lung sections then calculated as "Number of GFP+ cancer cells in lung". For inhibitor experiment, DMSO control or SBI-581 (30mg/kg) was injected intraperitoneally, then 30 minutes later GFP-labeled C8161.9 cells were injected through tail vein.
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