Gene editing of α6 integrin inhibits muscle invasive networks and increases cell-cell biophysical properties in prostate cancer

Cynthia S. Rubenstein, Jaime M.C. Gard, Mengdie Wang, Julie E. McGrath, Nadia Ingabire, James P. Hinton, Kendra D. Marr, Skyler J. Simpson, Raymond B Nagle, Cindy K. Miranti, Noel A. Warfel, Joe G.N. Garcia, Hina Arif-Tiwari, Anne E Cress

Research output: Contribution to journalArticle

Abstract

Human prostate cancer confined to the gland is indolent (low-risk), but tumors outside the capsule are aggressive (high-risk). Extracapsular extension requires invasion within and through a smooth muscle-structured environment. Because integrins respond to biomechanical cues, we used a gene editing approach to determine if a specific region of laminin-binding α6β1 integrin was required for smooth muscle invasion both in vitro and in vivo. Human tissue specimens showed prostate cancer invasion through smooth muscle and tumor coexpression of α6 integrin and E-cadherin in a cell-cell location and α6 integrin in a cell-extracellular matrix (ECM) distribution. Prostate cancer cells expressing α6 integrin (DU145 α6WT) produced a 3D invasive network on laminin- containing Matrigel and invaded into smooth muscle both in vitro and in vivo. In contrast, cells without α6 integrin (DU145 α6KO) and cells expressing an integrin mutant (DU145 α6AA) did not produce invasive networks, could not invade muscle both in vitro and in vivo, and surprisingly formed 3D cohesive clusters. Using electric cell-substrate impedance testing, cohesive clusters had up to a 30-fold increase in normalized resistance at 400 Hz (cell-cell impedance) as compared with the DU145 α6WT cells. In contrast, measurements at 40,000 Hz (cell-ECM coverage) showed that DU145 α6AA cells were two-fold decreased in normalized resistance and were defective in restoring resistance after a 1 μmol/L S1P challenge as compared with the DU145α6WTcells. The results suggest that gene editing of a specific α6 integrin extracellular region, not required for normal tissue function, can generate a new biophysical cancer phenotype unable to invade the muscle, presenting a new therapeutic strategy for metastasis prevention in prostate cancer. Significance: This study shows an innovative strategy to block prostate cancer metastasis and invasion in the muscle through gene editing of a specific α6 integrin extracellular region.

Original languageEnglish (US)
Pages (from-to)4703-4714
Number of pages12
JournalCancer Research
Volume79
Issue number18
DOIs
StatePublished - Sep 15 2019

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Integrins
Prostatic Neoplasms
Muscles
Smooth Muscle
Laminin
Electric Impedance
Extracellular Matrix
Gene Editing
Smooth Muscle Tumor
Neoplasm Metastasis
Cadherins
Capsules
Cues
Neoplasms
Phenotype

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

Cite this

Gene editing of α6 integrin inhibits muscle invasive networks and increases cell-cell biophysical properties in prostate cancer. / Rubenstein, Cynthia S.; Gard, Jaime M.C.; Wang, Mengdie; McGrath, Julie E.; Ingabire, Nadia; Hinton, James P.; Marr, Kendra D.; Simpson, Skyler J.; Nagle, Raymond B; Miranti, Cindy K.; Warfel, Noel A.; Garcia, Joe G.N.; Arif-Tiwari, Hina; Cress, Anne E.

In: Cancer Research, Vol. 79, No. 18, 15.09.2019, p. 4703-4714.

Research output: Contribution to journalArticle

Rubenstein, CS, Gard, JMC, Wang, M, McGrath, JE, Ingabire, N, Hinton, JP, Marr, KD, Simpson, SJ, Nagle, RB, Miranti, CK, Warfel, NA, Garcia, JGN, Arif-Tiwari, H & Cress, AE 2019, 'Gene editing of α6 integrin inhibits muscle invasive networks and increases cell-cell biophysical properties in prostate cancer', Cancer Research, vol. 79, no. 18, pp. 4703-4714. https://doi.org/10.1158/0008-5472.CAN-19-0868
Rubenstein, Cynthia S. ; Gard, Jaime M.C. ; Wang, Mengdie ; McGrath, Julie E. ; Ingabire, Nadia ; Hinton, James P. ; Marr, Kendra D. ; Simpson, Skyler J. ; Nagle, Raymond B ; Miranti, Cindy K. ; Warfel, Noel A. ; Garcia, Joe G.N. ; Arif-Tiwari, Hina ; Cress, Anne E. / Gene editing of α6 integrin inhibits muscle invasive networks and increases cell-cell biophysical properties in prostate cancer. In: Cancer Research. 2019 ; Vol. 79, No. 18. pp. 4703-4714.
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abstract = "Human prostate cancer confined to the gland is indolent (low-risk), but tumors outside the capsule are aggressive (high-risk). Extracapsular extension requires invasion within and through a smooth muscle-structured environment. Because integrins respond to biomechanical cues, we used a gene editing approach to determine if a specific region of laminin-binding α6β1 integrin was required for smooth muscle invasion both in vitro and in vivo. Human tissue specimens showed prostate cancer invasion through smooth muscle and tumor coexpression of α6 integrin and E-cadherin in a cell-cell location and α6 integrin in a cell-extracellular matrix (ECM) distribution. Prostate cancer cells expressing α6 integrin (DU145 α6WT) produced a 3D invasive network on laminin- containing Matrigel and invaded into smooth muscle both in vitro and in vivo. In contrast, cells without α6 integrin (DU145 α6KO) and cells expressing an integrin mutant (DU145 α6AA) did not produce invasive networks, could not invade muscle both in vitro and in vivo, and surprisingly formed 3D cohesive clusters. Using electric cell-substrate impedance testing, cohesive clusters had up to a 30-fold increase in normalized resistance at 400 Hz (cell-cell impedance) as compared with the DU145 α6WT cells. In contrast, measurements at 40,000 Hz (cell-ECM coverage) showed that DU145 α6AA cells were two-fold decreased in normalized resistance and were defective in restoring resistance after a 1 μmol/L S1P challenge as compared with the DU145α6WTcells. The results suggest that gene editing of a specific α6 integrin extracellular region, not required for normal tissue function, can generate a new biophysical cancer phenotype unable to invade the muscle, presenting a new therapeutic strategy for metastasis prevention in prostate cancer. Significance: This study shows an innovative strategy to block prostate cancer metastasis and invasion in the muscle through gene editing of a specific α6 integrin extracellular region.",
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AU - Rubenstein, Cynthia S.

AU - Gard, Jaime M.C.

AU - Wang, Mengdie

AU - McGrath, Julie E.

AU - Ingabire, Nadia

AU - Hinton, James P.

AU - Marr, Kendra D.

AU - Simpson, Skyler J.

AU - Nagle, Raymond B

AU - Miranti, Cindy K.

AU - Warfel, Noel A.

AU - Garcia, Joe G.N.

AU - Arif-Tiwari, Hina

AU - Cress, Anne E

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