GPR30-Expressing Gastric Chief Cells Do Not Dedifferentiate But Are Eliminated via PDK-Dependent Cell Competition During Development of Metaplasia.
GPR30-Expressing Gastric Chief Cells Do Not Dedifferentiate But Are Eliminated via PDK-Dependent Cell Competition During Development of Metaplasia.
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DOI：
10.1053/j.gastro.2020.01.046
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发表时间：
2020-05
期刊：
影响因子：
29.4
通讯作者：
Koike K
中科院分区：
文献类型：
--
作者：
Hata M;Kinoshita H;Hayakawa Y;Konishi M;Tsuboi M;Oya Y;Kurokawa K;Hayata Y;Nakagawa H;Tateishi K;Fujiwara H;Hirata Y;Worthley DL;Muranishi Y;Furukawa T;Kon S;Tomita H;Wang TC;Koike K
胃主细胞是一种分泌消化酶的成熟细胞类型，通过去分化或转分化，被认为是化生和癌症的起源。然而，支持这一说法的研究存在技术限制，包括主要细胞标志物的特异性和所用药物的毒性问题。因此，我们试图识别在主细胞中特异表达的基因，并建立一个追踪这些细胞的模型。我们对Mist1-CreERT追踪的细胞进行了转录组分析，有或没有主细胞耗尽。将Gpr30-RTTA小鼠与Teto-Cre小鼠杂交，与R26-TdTomato小鼠杂交后进行谱系追踪。使用Mist1-CreERT、Kitl-CreERT、TFF1-Cre和Tff2-Cre小鼠与报告小鼠杂交，进行了额外的谱系追踪实验。小鼠被给予大剂量的他莫昔芬或DMP-77，或感染幽门螺杆菌，以诱导胃化生。我们使用Teto-KrasG12D、LSL-KrasG12D和LSL-HrasG12V小鼠研究了在胃细胞中表达突变形式的RAS的小鼠。我们分析了GPR30基因敲除小鼠的胃组织。给小鼠注射二氯乙酸酯以抑制依赖于丙酮酸脱氢酶激酶(PDK)的细胞竞争。我们发现GPR30是雌激素受体的G蛋白偶联形式，是小鼠胃上皮主细胞的细胞特异性标志物。Gpr30-RTTA小鼠与Teto-Cre杂交；R26-TdTomato小鼠在主细胞中有GPR30的特异性表达，在峡部干细胞和腺体的谱系追踪中没有表达。突变型Kras在GPR30+主细胞中的表达并未导致化生或异型增生的发展，而是导致主细胞标记数量的减少和来自于上Kitl+克隆的颈系的代偿性扩张。给予大剂量他莫昔芬、DMP-777或幽门螺杆菌可减少标记主细胞的数量。在化生刺激后，主要细胞通过依赖GPR30和PDK的细胞竞争从上皮细胞中消失，而GRP30的丢失或PDK活性的抑制保留了主要细胞的数量，并减弱了颈系细胞的扩张。在对小鼠的追踪研究中，我们发现大多数主要细胞在化生过程中丢失，因此不太可能与胃癌的发生有关。共同表达颈部和主要谱系标记的细胞的扩张，即所谓的痉挛多肽表达化生，不是通过主细胞的去分化发生的，而是通过颈部细胞的代偿反应来取代被消除的主细胞。
Gastric chief cells, a mature cell type that secretes digestive enzymes, have been proposed to be the origin of metaplasia and cancer, through dedifferentiation or trans-differentiation. However, studies supporting this claim have had technical limitations, including issues with the specificity of chief cell markers and the toxicity of drugs used. We therefore sought to identify genes expressed specifically in chief cells and establish a model to trace these cells. We performed transcriptome analysis of Mist1-CreERT-traced cells, with or without chief cell depletion. Gpr30-rtTA mice were generated, crossed to TetO-Cre mice, and lineage tracing was performed after crosses to R26-TdTomato mice. Additional lineage tracing experiments were performed using Mist1-CreERT, Kitl-CreERT, Tff1-Cre, and Tff2-Cre mice crossed to reporter mice. Mice were given high-dose tamoxifen or DMP-77, or infected with Helicobacter pylori, to induce gastric metaplasia. We studied mice that expressed mutant forms of Ras in gastric cells, using TetO-KrasG12D, LSL-KrasG12D, and LSL-HrasG12V mice. We analyzed stomach tissues from GPR30-knockout mice. Mice were given dichloroacetate to inhibit pyruvate dehydrogenase kinase (PDK)-dependent cell competition. We identified GPR30, the G-protein coupled form of the estrogen receptor, as a cell-specific marker of chief cells in gastric epithelium of mice. Gpr30-rtTA mice crossed to TetO-Cre; R26-TdTomato mice had specific expression of GPR30 in chief cells, with no expression noted in isthmus stem cells or lineage tracing of glands. Expression of mutant Kras in GPR30+ chief cells did not lead to the development of metaplasia or dysplasia, but instead led to a reduction in labeled numbers of chief cells and a compensatory expansion of neck lineage, which was derived from upper Kitl+ clones. Administration of high-dose tamoxifen, DMP-777, or H pylori decreased the number of labeled chief cells. Chief cells were eliminated from epithelia via GPR30- and PDK-dependent cell competition after metaplastic stimuli, whereas loss of GRP30 or inhibition of PDK activity preserved chief cell numbers and attenuated neck lineage cell expansion. In tracing studies of mice, we found that most chief cells are lost during metaplasia and therefore are unlikely to contribute to gastric carcinogenesis. Expansion of cells that co-express neck and chief lineage markers, known as spasmolytic polypeptide-expressing metaplasia, does not occur via dedifferentiation from chief cells, but rather through a compensatory response from neck cells to replace the eliminated chief cells.
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影响因子：
24.5
作者：
Hayakawa Y;Jin G;Wang H;Chen X;Westphalen CB;Asfaha S;Renz BW;Ariyama H;Dubeykovskaya ZA;Takemoto Y;Lee Y;Muley A;Tailor Y;Chen D;Muthupalani S;Fox JG;Shulkes A;Worthley DL;Takaishi S;Wang TC
通讯作者：
Wang TC
影响因子：
29.4
作者：
Huh WJ;Khurana SS;Geahlen JH;Kohli K;Waller RA;Mills JC
通讯作者：
Mills JC
DOI：
10.1016/j.jcmgh.2017.01.013
发表时间：
2017-05
影响因子：
7.2
作者：
Hayakawa Y;Fox JG;Wang TC
通讯作者：
Wang TC
DOI：
10.1126/science.1180794
发表时间：
2010-01-29
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Li L;Clevers H
通讯作者：
Clevers H
影响因子：
50.3
作者：
Hayakawa Y;Ariyama H;Stancikova J;Sakitani K;Asfaha S;Renz BW;Dubeykovskaya ZA;Shibata W;Wang H;Westphalen CB;Chen X;Takemoto Y;Kim W;Khurana SS;Tailor Y;Nagar K;Tomita H;Hara A;Sepulveda AR;Setlik W;Gershon MD;Saha S;Ding L;Shen Z;Fox JG;Friedman RA;Konieczny SF;Worthley DL;Korinek V;Wang TC
通讯作者：
Wang TC