| 研究課題/領域番号 |
23K06639
|
|---|
| 研究種目 |
基盤研究(C)
|
| 配分区分 | 基金 |
|---|
| 応募区分 | 一般 |
|---|
| 審査区分 |
小区分50010:腫瘍生物学関連
|
|---|
| 研究機関 | 慶應義塾大学 |
|---|
研究代表者 |
|
|---|
| 研究期間 (年度) |
2023-04-01 – 2026-03-31
|
| 研究課題ステータス |
交付 (2023年度)
|
| 配分額 *注記 |
4,030千円 (直接経費: 3,100千円、間接経費: 930千円)
2025年度: 1,170千円 (直接経費: 900千円、間接経費: 270千円)
2024年度: 1,560千円 (直接経費: 1,200千円、間接経費: 360千円)
2023年度: 1,300千円 (直接経費: 1,000千円、間接経費: 300千円)
|
|---|
| キーワード | metastasis / CXCR7 / cancer / chemokine / ACKR3 |
|---|
| 研究開始時の研究の概要 |
The proposed research aims to improve our understanding of the mechanisms regulating cancer metastasis, the leading cause for cancer death.
The focus is on the chemokine receptor ACKR3, recently identified from a screening for genes regulating metastasis.
Another goal is to develop methods for labeling, isolating and characterizing cells from the metastatic niche, the non-cancer cells surrounding the tumor.
Successful implementation of the research is expected to make scientific and potentially clinical contributions in the field of cancer metastasis.
|
| 研究実績の概要 |
Initial results with the CXCR7 neutralizing antibody X7Ab suggested suppression of metastatic growth of the colon cancer cell line 2017 transplanted in NOG mice when compared with the control Rituximab antibody.
However, the result was not doze dependent - the suppression effect was observed only with a doze of 2.4 mg/kg X7Ab and not with the higher doze of 5 mg/kg. A repetition of the experiment with 2.4 mg/kg X7Ab failed to replicate the previous results. Likewise, the CXCR7 inhibitor ACT-1004-1239 did not suppress metastatic growth of 2017. Detailed analysis suggested that the previously observed CXCR7 KO effects might be due to clonal differences in an intrinsically heterogeneous 2017 cell line. Moreover, RNAseq, real-time PCR and FACS sorting with X7Ab experiments demonstrated that 2017 displayed very low expression of CXCR7.
Currently I am replicating the CXCR7 KO results in a separate cell line, 2044, which I verified to be expressing high levels of CXCR7.
In vivo experiments with organoids expressing the in vivo labeling marker sLP-mCherry demonstrated stable expression of the marker, which however remained largely confined within the tumor and failed to label the surrounding host cells.
|
| 現在までの達成度 (区分) |
現在までの達成度 (区分)
3: やや遅れている
理由
The results with the cell line 2017 were not very stable, probably due to the low expression level of the target protein CXCR7 and strong clonal effects.
The in vivo label sLP-mCherry did not diffuse from the tumor cells to label the neighboring host cells.
|
| 今後の研究の推進方策 |
I verified high expression of CXCR7 in a separate cell line, 2044, isolated several clones from it and identified a clone with metastatic ability. I used this clone to knock out CXCR7, isolated and verified CXCR7 KO in several clones. I will proceed to compare the metastatic ability of the CXCR7 KO clones with the wild type clone. If the CXCR7 KO clones are indeed impaired in their metastatic ability, I will compare the RNA profiles of the CXCR7 KO with the wild type clone to identify potential pathways underlying the difference.
Recent advances in spatial transcriptomic analysis techniques such as Xenium and Visium from 10X Genomics can bypass the need for labeling and extracting cancer-neighboring cells and help me identify CXCR7-dependent gene expression changes in host cells.
|
