In Our View, the Frequency of Cells That Can Sustain Tumor Growth, and Thus the Generality

平成20年度

医学系研究科再生医科学専攻

組織・器官形成

外国語（組織・器官形成）

次の英文を読み、問いに答えよ

Cancer biologists are intrigued by the hypothesisthat tumor growth may be sustained by a rare subpopulation of the cells, termed cancer stem cells. Supporting this concept are the heterogeneous cellular composition of certain tumors and the finding that only a minute proportion of the cells (~1/106) in some human acute myeloid leukemia (AML) samples can seed tumor growth when transplanted into sublethally irradiated nonobese diabetic (NOD) severe combined immunodeficient (scid) mice. The interpretation of such xenotransplantation studies, however, is complicated by the critical role in tumor growth of interactions with the microenvironment, which are mediated by both soluble and membrane-bound factors. Notably, many such mouse factors cannot engage the cognate human receptor and vice versa. ①Thus, the low frequency of human AML cells producing tumors in NOD/scid mice might reflect in part the rarity of human tumor cells that can readily adapt to growth in a foreign (mouse) milieu.

②In our view, the frequency of cells that can sustain tumor growth, and thus the generality of the cancer stem cell hypothesis, can best be tested by transfer of titrated numbers of mouse tumor cells into nonirradiated histocompatible recipient mice. We isolated primary pre-B/B lymphoma cells from three independent Em-myc transgenic mice and injected 10 to 105cells into nonirradiated congenic animals. Regardless of the cell number injected, all recipients became moribund with disseminated lymphoma within 35 days (Table 1). Although the number of injected cells did not noticeably affect tumor burden, organ infiltration, or disease severity, recipients of 10 or 100 lymphoma cells usually developed tumors more slowly than those receiving 105cells. ③Importantly, even transfer of a single cell elicited fatal lymphoma in three of eight recipients within 33 to 76 days (case 2).

A small fraction (~2 to 5%) of the cells in primary Em-myc lymphomas displayed the characteristic stem cell markers Sca-1 and/or AA4.1. However, when sorted Sca-1+ AA4.1hi or Sca-1+ AA4.1lo lymphoma cells were transplanted, as few as 10 cells of each population elicited fatal lymphoma within 17 to 40 days (Table 1). Similarly, with Em-N-RAS thymic lymphomas and four independent cases of AML caused by PU.1 deficiency, recipients transplanted with as few as 10 cells developed tumors, although onset was delayed in mice receiving only 10 or 100 AML cells (Table 1). For all three malignancies, the cell surface marker phenotype , the gene expression profile , and the invasiveness of the transplanted tumors mirrored that of the primary tumor.

These observations challenge the concept that growth of AML, and possibly other malignancies, are always sustained by a rare cancer stem cell. Although cancer stem cells may well drive the growth of many cancers, particularly those displaying extensive differentiation, our studies of mouse lymphomas and leukemias indicate that at least certain malignancies (particularly those with substantial homogeneity) can be maintained by a relatively large proportion (>10%) of tumor cells, perhaps even the majority. ④Although mouse and human tumors differ in notable respects, the marked disparity with results from human AML cells suggests that xenotransplantation may underestimate the percentage of tumor-sustaining cells. With common human solid tumors (for example, brain, colon, and breast), transplantation places the tumor growth–sustaining cells within subpopulations (for example, CD133+) that compose up to 20% of the cells, and most of the remaining cells might be at differentiation stages unsupportable by the mouse microenvironment. The reported rarity of cancer stem cells in AML and colon cancer might reflect the need to cotransfer an essential human accessory cell (we note that endothelial cell progenitors are also CD133+).

⑤Determining whether the growth of various tumors is sustained by most of the tumor cells or by a rare subpopulation has important ramifications for the design of novel therapies. Therefore, the cancer stem cell hypothesis merits more rigorous tests. ⑥For human tumors, ultimately this will require transfer of tumor cells into mice installed with all the requisite human support cells. Lastly, because the term “cancer stem cell” also currently designates the normal cell that founded the tumor, we suggest that the cells sustaining growth of an established tumor be referred to as “⑦tumor-propagating cells.”

Table 1. A large proportion of tumor cells can sustain the growth of murine lymphoid and myeloid malignancies. Cells from primary Em-myc pre-B/B lymphomas, Em-N-RAS thymic lymphomas, or PU.1−/− AML, all from mice on a C57BL/6 (Ly5.2+) background (>15 backcrosses), were transplanted into nonirradiated congenic C57BL/6 (Ly5.1+) recipient mice. To circumvent problems associated withinjection of low cell numbers, we mixed the tumor cells with 106congenic (C57BL/6-Ly5.1+) spleen cells as carriers. Shown are the fraction of recipients that developed tumors and the average time from transplantation to tumor development. No mice (0/24) injected with carrier spleen cells alone developed any tumor over a 100-day period. ND, not determined.

Recipients that developed tumors (days to kill)

105cells 103cells 102cells 10 cells

Em-myc B lymphoma

Case 1 3/3 (25) 3/3 (25) 3/3 (32) 2/2 (35)

Case 2 3/3 (21) 3/3 (23) 3/3 (24) 3/3 (24)

Case 3 Sca-1+ AA4.1hi 3/3 (21) 3/3 (21) ND 3/3 (17)

Sca-1+ AA4.1lo 2/2 (17) 2/2 (28) 2/2 (28) 2/2 (40)

Em-N-RAS T lymphoma

Case 1 3/3 (28) 3/3 (42) 3/3 (28) 3/3 (28)

PU.1–/– AML

Case 1 1/1 (54) 2/2 (168) 1/2 (192) 0/2

Case 2 2/2 (84) 2/2 (85) 2/2 (224) 1/2 (114)

Case 3 1/1 (85) 2/2 (62) 2/2 (69) 2/2 (90)

Case 4 1/1 (30) 1/1 (37) 2/2 (79) 2/2 (88)

Science 317, 337, 2007より一部改変して引用

(参考)

sublethally irradiated nonobese diabetic (NOD) severe combined immunodeficient (scid) mice: 亜致死量放射線照射された非肥満性糖尿病重症複合性免疫不全マウス。免疫担当細胞のほとんどを失わせたマウスで、異種の細胞が効率よく長期間にわたって移植可能になる。

Histocompatible：組織を移植しても拒絶反応が起こらない。

the cell surface marker phenotype：細胞表面マーカーで判別された表現型。細胞表面の蛋白質（本文中のSca-1など）の発現パターンにより細胞を分類すること。

the gene expression profile：遺伝子発現プロフィール。DNAアレーなどを用いて、細胞の分類すること。

問１

下線①を訳せ

問２

下線②を訳せ。

問３

下線②の実験内容の概略を簡潔に延べよ。Table1の説明を参考にして良い。

問４

下線③について、transfer of a single cell がfatal lymphoma をthree of eight recipients に発症させることがなぜ従来のcancer stem cellの考えに再考を迫ることになるのか、わかりやすく説明せよ。

問５

下線④を訳せ

問６

下線④のtumor-sustaining cellsの実例と予想される細胞を一つ挙げよ

問７

下線⑤について、growth of various tumors が most of the tumor cells によって維持されるのかあるいはa rare subpopulation によって維持されるのかを決めることが、ガンの新療法の開発になぜ重要なのか、あなたが一つの新しい抗ガン剤を開発していると仮定して、簡潔に述べよ。

問８

下線⑥を訳せ。

問９

下線⑦のtumor-propagating cellsとtumor-sustaining cellsは同じ細胞を示すのか、違う細胞を示すのか、答えよ。

問１０

Table1について、PU.1–/– AMLマウスのAML細胞を移植したときの実験結果を、case3についてのみ文章で説明せよ。

問１１

本論文のタイトルを考えよ。答えは英語あるいは日本語で示せ。