Construction of PDX library derived from Japanese cancer patients○ Shinichiro Tsunesumi1, Yukiko Shimada2, Shigenori Enoki1, Seiichi Katayama1, Shigehiro Yagishita3, Mami Takahashi3,33
Toshio Imai , Akinobu Hamada
1: LSIM Safety Institute Corporation, 2: LSI Medience Corporation, 3: National Cancer Center Research Institute

Summary in JapaneseMaterials and Methods

抗がん剤開発の成功率を向上させるためには，臨床的に適切な腫瘍モデルが求められている．ヒト腫瘍を免疫不全マウスに直接移植することによって作製される患者由来の異種移植片（PDX）モデルは，従来のモデルよりも臨床的に適切なモデルとして広く受け入れられている．我々は，日本人の特性も加味した効率的な新薬開発を目指して，日本人がん患者由来の PDX（J-PDX）を産業活用できるよう，またさらに高度な研究・医療への応用の推進に取り組んできた．

AnimalFemale NOG mice (NOD.Cg-Prkdc scidIl2rg tm1Sug/ShiJic) obtained from Central Institute for Experimental Animals.

PDX tumor

PDX tumors obtained from NCC and stored in a liquid nitrogen storage tanks.

Inoculation of PDX tumor

国立がん研究センターより入手した PDX 腫瘍を免疫不全マウスに移植した．一定期間後に腫瘍を採取し，腫瘍片を液体窒素タンクに保存し，ライブラリーとして整備した．また，採取した腫瘍の一部は病理解析や遺伝子解析を行い，各 PDX の情報を収集したため，そのデータの一部を紹介する．

PDX tumor pieces were inoculated subcutaneously or into the mammary gland under isoﬂurane inhalation
anesthesia. PDX tumors were inoculated to 3 animals per strain.

Measurement of tumor diameters

これらの情報を整備することは，抗がん剤の開発に使用する PDX の選択に非常に重要であり，新薬の開発に大いに貢献するものである．

Tumor diameter was measured using caliper once a week. The tumor volume was calculated by the followingequation.
Estimated tumor volume (mm3) = 1/2 × long diameter (mm) × short diameter (mm) × short diameter (mm)Pathological analysis

Objective

Tumor was immersed in 10% formalin neutral buﬀer solution, and paraﬃn-embedded blocks were created.
Pathological analysis (histological type) was performed using HE stained tissue sections at Pathology Institute Corp.

Whole exome sequencing

To improve the success rate of anti-cancer drug development, clinically relevant tumor
models are required. Patient-derived xenograft (PDX) tumor model, which is generated bydirect implantation of human tumor into immunodeﬁcient mice, has been widely accepted
as a more clinically relevant model than conventional model. We have been promoting theapplication of PDX derived from Japanese cancer patients to more advanced research andmedical treatment so that it can be industrially utilized with the aim of developing eﬃcientnew drugs that take into account the characteristics of Japanese people. PDX tumors
obtained from the National Cancer Center (NCC) were transplanted into immunodeﬁcient
mice in the LSIM Safety Institute Corporation (LSSI). Tumors were collected and the tumor
pieces were stored in liquid nitrogen tanks and maintained as libraries. In addition, some of
the collected tumors were subjected to pathological analysis and gene analysis to collectinformation on each PDX.

Results

Tumor was snap-frozen, and RNA was extracted using AllPrep DNA/RNA 96 Kit (QIAGEN).
Whole exome sequencing was performed at Macrogen Japan Corp., and analyzed by the Genetic Analysis ResearchDepartment, LSI Medience Corp.

Conclusion
There was a large diﬀerence in the growth rate between tumors. Even in tumor pieces of thesame PDX, the growth rate was diﬀerent for each animal. Compared to the CDX model inwhich a homogeneous cell population is inoculated, the PDX model, which maintains cancer
heterogeneity, tends to have greater variation in tumor growth. The present data are usefulfor pharmacological studies using PDXs because this variation should be taken into accountwhen setting the number of animals in the group, and the number of inoculated animals forthe required number of animals.

The red, blue, and green lines in each graph show the growth of tumors in each animal.

800

1200

400

10002002W

4W

6W

8W 10W 12W 14W 16W

Weeks after inoculation

LSIM ID:
BI-001-LSIM
Gene mutation: CDK13600
40020000W

2W

4W

6W

158W 10W 12W 14W 16W

Weeks after inoculation

1313

LSIM ID:
PA-004-LSIM
Gene mutation: KRAS, POLH

1200

1200

1000

1000

800
600
400200010 110W

2W

4W

6W

80060040020008W 10W 12W 14W 16W

Weeks after inoculation

LSIM ID:
CO-005-LSIM
Gene mutation: KRAS, TP53

107

150W

2W

4W

6W

8W 10W 12W 14W 16W

Weeks after inoculation

LSIM ID:
CO-031-LSIM
Gene mutation: ALPL, CDKL5, PIK3CA, etc.

Colorectalcancer

18
1926

Lung
cancerBreast cancer
1200

99

Number of PDXs by cancer type
410 PDX tumors were established by NCC.
153 PDX tumors have been preserved after growth in LSSI.

Rare cancers, Sarcoma

Lung cancer

600
4002000W

2W

4W

6W

8W 10W 12W 14W 16W

Weeks after inoculation

LSIM ID:
BC-005-LSIM
Gene mutation: TP53

120012001000

1000

80060040020000W

2W

4W

6W

8W 10W 12W 14W 16W

Weeks after inoculation

LSIM ID:
LU-015-LSIM
Gene mutation: C4B, EGFR, TNXB

1200

8006004002000Ewing Sarcoma

1000800600
40020000W

2W

4W

6W

8W 10W 12W 14W 16W

Weeks after inoculation

LSIM ID:
LU-022-LSIM
Gene mutation: KMT2D, NFE2L2, PKHD10W

2W

4W

6W

8W 10W 12W 14W 16W

Weeks after inoculation

LSIM ID:
RA-005-LSIM
Gene mutation: PRNP

Please refer to our website for PDX list, gene expression information, details of gene mutation, etc.
https://www.ankaken.co.jp/topics02.html

Rhabdomyosarcoma

1200

Tumor volume (mm3)800Tumor volume (mm3)

1000

0

Rare cancers,
Sarcoma64
Tumor volume (mm3)

0W

800

Tumor volume (mm3)0Tumor volume (mm3)600Tumor volume (mm3)

Pancreatic cancer

1000

Tumor volume (mm3)Tumor volume (mm3)1200Colorectal cancer
Tumor volume (mm3)

Biliary tract cancer

Endometrial cancer
Brain tumor
Gastric cancer
Ovarian cancer
Esophageal cancer
Other

1000
800
600
400
20000W

2W4W

6W

8W 10W 12W 14W 16W

Weeks after inoculation

LSIM ID:
RA-016-LSIM
Gene mutation: GALC, GJB2COI Disclosure Information
Lead Presenter: Shinichiro Tsunesumi
I have no ﬁnancial relationships to disclose.