{"id":416,"date":"2022-06-01T23:39:49","date_gmt":"2022-06-01T23:39:49","guid":{"rendered":"https:\/\/students.pingry.org\/irt\/?page_id=416"},"modified":"2023-03-31T12:37:52","modified_gmt":"2023-03-31T12:37:52","slug":"kif-yap-2022-project","status":"publish","type":"page","link":"https:\/\/students.pingry.org\/irt\/kif-yap-2022-project\/","title":{"rendered":"Kif\/Yap – 2022 Project"},"content":{"rendered":"

[et_pb_section fb_built=”1″ admin_label=”Hero” _builder_version=”4.4.7″ background_color=”#f8fbfe” use_background_color_gradient=”on” background_color_gradient_start=”#53a0fd” background_color_gradient_end=”#002960″ background_color_gradient_direction=”74deg” background_image=”https:\/\/students.pingry.org\/irt\/wp-content\/uploads\/sites\/19\/2020\/07\/geometric-bg-overlay-01.jpg” background_blend=”overlay” custom_padding=”2.4%|0px||0px||” animation_style=”zoom” animation_direction=”bottom” animation_duration=”0ms” animation_intensity_zoom=”0%” animation_starting_opacity=”100%” box_shadow_style=”preset7″ box_shadow_horizontal=”0px” box_shadow_vertical=”-200px” box_shadow_color=”#F8FBFE”][et_pb_row _builder_version=”4.4.7″ custom_padding=”||0px|||”][et_pb_column type=”4_4″ _builder_version=”3.25″ custom_padding=”|||” custom_padding__hover=”|||”][et_pb_text _builder_version=”4.4.7″ text_font=”Abhaya Libre||||||||” text_text_color=”#ffffff” text_font_size=”18px” text_line_height=”1.8em” header_font=”Nunito Sans|700|||||||” header_text_align=”center” header_text_color=”#ffffff” header_font_size=”45px” header_line_height=”1.3em” header_2_font=”||||||||” text_orientation=”center” max_width=”700px” module_alignment=”center” custom_margin=”||18px|||” custom_padding=”||17px|||” inline_fonts=”Abhaya Libre”]<\/p>\n

Kif\/Yap 2021\/2022 Project –\u00a0The Effect of KIF11 Activity on YAP Localization<\/span><\/h1>\n

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Luca Pizzale (Form VI), Elodie Wardle (Form VI), \u00a0John Grissinger (Form V),
Matt Oatman (Form V), \u00a0Katia Krishtopa (Form IV)<\/span><\/h3>\n

[\/et_pb_text][et_pb_image align=”center” align_tablet=”center” align_phone=”” align_last_edited=”on|desktop” _builder_version=”4.4.7″ min_height=”26px” custom_margin=”41px||-79px|||” custom_padding=”||0px|||” border_radii=”on|10px|10px|10px|10px” border_width_all=”1px” border_color_all=”rgba(51,51,51,0.1)” box_shadow_style=”preset1″ box_shadow_vertical=”10px” box_shadow_blur=”30px” box_shadow_color=”rgba(34,56,101,0.09)”][\/et_pb_image][\/et_pb_column][\/et_pb_row][et_pb_row _builder_version=”4.4.7″][et_pb_column type=”4_4″ _builder_version=”4.4.7″][et_pb_text _builder_version=”4.4.7″ text_font=”Abhaya Libre|600|||||||” header_text_align=”center” header_font_size=”54px” min_height=”86px” custom_margin=”||-9px|||”]<\/p>\n

Project Summary + Project Goal\u00a0<\/h1>\n

[\/et_pb_text][\/et_pb_column][\/et_pb_row][\/et_pb_section][et_pb_section fb_built=”1″ admin_label=”Features” _builder_version=”4.4.7″ background_color=”#f8fbfe” min_height=”2490px” custom_margin=”-62px|||||” custom_padding=”151px|0px|12px|0px||”][et_pb_row _builder_version=”4.4.7″ min_height=”132px” custom_margin=”-10px|auto||auto||” custom_padding=”||0px|||”][et_pb_column type=”4_4″ _builder_version=”4.4.7″][et_pb_text _builder_version=”4.4.7″ text_font=”Abhaya Libre||||||||” text_text_color=”#ffffff” text_line_height=”1.9em” background_color=”#cee1ff” width=”100%” max_width=”100%” min_height=”382px” max_height=”1000px” custom_margin=”-190px|-1px|0px|-2px|false|false” custom_padding=”14px||1px|4px|false|false” border_width_all=”5px” border_style_all=”inset”]<\/p>\n

This project is founded upon targeted gene\u00a0therapy and two potential\u00a0proteins\u00a0involved: KIF11 and YAP. KIF11, a motor\u00a0protein, and YAP, a\u00a0transcription factor are both potential drivers of cancer and possibly may be\u00a0related to one another. Investigating these two\u00a0drivers may provide\u00a0helpful\u00a0information in future targeted\u00a0therapies which is why the group chose to\u00a0focus\u00a0specially on their\u00a0unknown\u00a0connection.\u00a0<\/span><\/h2>\n

Upon consulting Dr. Gao at Rutgers\u00a0Laboratory, the team hypothesized that KIF11\u00a0participates in the cellular transportation of YAP (in other words: KIF11 and YAP are connected). It was known that when active, YAP lives in the\u00a0<\/span>nucleus and when inactive, YAP is phosphorilated in the cytoplasm. From this past knowledge, the group developed a \u00a0potential pathway to show KIF11 and YAP connections (Figure 1). Both protiens might be connected through Rab11a, a GTPase that regulates intercellular traffic. Rab11a has an inverse relationship with YAP (the less Rab11a leads to more activated YAP). Dr. Gao found that KIF11 was found to bind to Rab11a when intact which supported this pathway (Figure 2).<\/h2>\n

From our pathway, the group then focused on the question “If we inhibit KIF11 activity, does YAP’s location change”. The original hypothesis to answer this question is: as KIF11 is inhibited, YAP becomes inactive and moves to the cytoplasm. To do this, the group used SW480 cells (human colon cancer cells) provided by Dr. Gao, and grew, harvested, and froze them for experimentation. To inhibit KIF11, 6 plates were treated with ispinesib (concentrations ranging from 0 to 1000 nM) for 24 hours and then the contents of each plate were frozen and thawed to break down all cells. To separate the nuclear and cytosolic fragments of each cell, a centrifuge was used and then the cell contents were ran through a western blot test (Figure 3).\u00a0<\/h2>\n

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Current State<\/h2>\n

[\/et_pb_text][\/et_pb_column][\/et_pb_row][et_pb_row _builder_version=”4.4.7″ min_height=”267px” custom_margin=”-68px|auto||auto||” custom_padding=”41px||0px|||”][et_pb_column type=”4_4″ _builder_version=”4.4.7″][et_pb_text _builder_version=”4.4.7″ text_font=”Abhaya Libre||||||||” header_letter_spacing=”1px” background_color=”#cee1ff” min_height=”178px” custom_margin=”||48px|||” custom_padding=”15px||8px|4px|false|false” border_width_all=”5px” border_style_all=”inset”]<\/p>\n

Our team has ran our final western\u00a0<\/span>blot to explore and confirm our hypothesis (Figure 3). This blot has shown us the presence of KIF11 and YAP as well as how the inhibition of KIF11 appears to change the location of YAP from the nucleus to the cytoplasm. The blot showed that as the concentration of Ispinesib increases, the protein band representing the nuclear YAP fades, indicating that the amount of YAP in the nucleus is decreasing. As it is known that YAP is in the nucleus when it is dephosphorylated and active, but it is in the cytoplasm when it is phosphorylated and inactive and the western suggests that KIF11 inhibition\u00a0<\/span>casques<\/span>\u00a0YAP to become\u00a0<\/span><\/span><\/span>phosphorylated and exit the\u00a0nucleus, therefore becoming inactive. These\u00a0results\u00a0support the hypothesis that there is a connection between the proteins.\u00a0<\/span><\/span>Although the\u00a0<\/span>group<\/span>\u00a0had some\u00a0staining difficulties on the last western blot (where the same\u00a0proceidgers were followed) (Figure 4), what was\u00a0visible\u00a0coordinated directly with our\u00a0previous blot which supported our past findings. Both blots have\u00a0answered how KIF11 and YAP are connected as well as the location of both\u00a0proteins when one is\u00a0inhibited or\u00a0activated. Due to time\u00a0constraints, the group could not\u00a0repeat our experiment once more or change the control\u00a0variable to test\u00a0accuracy,\u00a0however the group is\u00a0confident in our\u00a0findings.\u00a0<\/span><\/span><\/span><\/p>\n

As we have proved our hypothesis, our project has concluded and will no longer be pursued in the upcoming years.\u00a0<\/span><\/p>\n

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Next Steps<\/span><\/h2>\n

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Our final goal is to share our findings though the Pingry\u00a0Community Research\u00a0Journal as we have concluded our project. We hope to publish the connection we made between KIF11 and YAP so that\u00a0further research may be\u00a0done on\u00a0both\u00a0proteins in the future.\u00a0<\/span><\/h2>\n

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Research Week Poster\u00a0<\/span><\/h2>\n

[\/et_pb_text][\/et_pb_column][\/et_pb_row][et_pb_row _builder_version=”4.4.7″ custom_margin=”|auto|-38px|auto||”][et_pb_column type=”4_4″ _builder_version=”4.4.7″][et_pb_image src=”https:\/\/students.pingry.org\/irt\/wp-content\/uploads\/sites\/19\/2022\/06\/Copy-of-2022-KIF_YAP-poster.jpg” title_text=”Copy of 2022 KIF_YAP poster” _builder_version=”4.4.7″ custom_margin=”-39px|||||” border_width_all=”4px” border_style_all=”inset”][\/et_pb_image][\/et_pb_column][\/et_pb_row][\/et_pb_section][et_pb_section fb_built=”1″ _builder_version=”4.4.7″][et_pb_row _builder_version=”4.4.7″][et_pb_column type=”4_4″ _builder_version=”4.4.7″][et_pb_text _builder_version=”4.4.7″ header_font=”Abhaya Libre|600|||||||” header_2_font=”Abhaya Libre|600|||||||” header_2_font_size=”54px” custom_margin=”-41px|||||” inline_fonts=”Abhaya Libre”]<\/p>\n

References<\/span><\/h2>\n

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Purcell, J. W. et al. (2010). Activity of the kinesin spindle protein inhibitor\u00a0ispinesib (SB-715992) in models of breast cancer. Clinical Cancer Research<\/i>, 16<\/i>(2), 566-576.\u00a0<\/h2>\n

D’Agostino, L. (2019). Recycling endosomes in mature epithelia restrain\u00a0tumorigenic signaling. Cancer Research<\/i>, 79<\/i>(16), 4099-4112.<\/h2>\n

[\/et_pb_text][\/et_pb_column][\/et_pb_row][\/et_pb_section][et_pb_section fb_built=”1″ admin_label=”Center Aligned Text ” _builder_version=”4.4.7″ background_color=”#f8fbfe” use_background_color_gradient=”on” background_color_gradient_start=”#53a0fd” background_color_gradient_end=”#002960″ background_color_gradient_direction=”74deg” background_image=”https:\/\/students.pingry.org\/irt\/wp-content\/uploads\/sites\/19\/2020\/07\/geometric-bg-overlay-01.jpg” background_blend=”overlay” min_height=”223px” custom_margin=”-28px||-330px|||” custom_padding=”0px|0px|0px|0px||” animation_direction=”bottom”][et_pb_row admin_label=”Tittle” _builder_version=”4.4.7″ custom_margin=”1px|auto||auto||”][et_pb_column type=”4_4″ _builder_version=”3.25″ custom_padding=”|||” custom_padding__hover=”|||”][et_pb_text _builder_version=”4.4.7″ header_font=”Abhaya Libre|700|||||||” header_text_align=”center” header_text_color=”#ffffff” header_font_size=”60px” custom_padding=”||0px|||”]<\/p>\n

Meet the Team<\/h1>\n

[\/et_pb_text][\/et_pb_column][\/et_pb_row][et_pb_row column_structure=”1_5,1_5,1_5,1_5,1_5″ _builder_version=”4.4.7″ min_height=”85px” height=”87px” max_height=”100px” custom_margin=”||-2px||false|false” custom_padding=”0px||0px||false|false”][et_pb_column type=”1_5″ _builder_version=”4.4.7″][et_pb_image src=”https:\/\/students.pingry.org\/irt\/wp-content\/uploads\/sites\/19\/2020\/08\/Pizzale_Luca_Grade-10_2019-20.jpg” title_text=”Pizzale_Luca_Grade 10_2019-20″ _builder_version=”4.4.7″ transform_scale=”118%|100%” transform_scale_linked=”off” transform_translate=”9px|0px” transform_translate_linked=”off” width=”94%” height=”211px”][\/et_pb_image][\/et_pb_column][et_pb_column type=”1_5″ _builder_version=”4.4.7″][et_pb_image src=”https:\/\/students.pingry.org\/irt\/wp-content\/uploads\/sites\/19\/2022\/06\/G12-Wardle_Elodie.jpg” title_text=”G12 Wardle_Elodie” _builder_version=”4.4.7″][\/et_pb_image][\/et_pb_column][et_pb_column type=”1_5″ _builder_version=”4.4.7″][et_pb_image src=”https:\/\/students.pingry.org\/irt\/wp-content\/uploads\/sites\/19\/2022\/06\/G11-Grissinger-J.jpg” title_text=”G11 Grissinger J” _builder_version=”4.4.7″ min_height=”213px”][\/et_pb_image][\/et_pb_column][et_pb_column type=”1_5″ _builder_version=”4.4.7″][et_pb_image src=”https:\/\/students.pingry.org\/irt\/wp-content\/uploads\/sites\/19\/2022\/06\/G11-Oatman-M.jpg” title_text=”G11 Oatman M” _builder_version=”4.4.7″ height=”213px”][\/et_pb_image][\/et_pb_column][et_pb_column type=”1_5″ _builder_version=”4.4.7″][et_pb_image src=”https:\/\/students.pingry.org\/irt\/wp-content\/uploads\/sites\/19\/2023\/03\/zje1gp6ritzriagpkzzt.jpeg” title_text=”zje1gp6ritzriagpkzzt” _builder_version=”4.4.7″ height=”214px” hover_enabled=”0″][\/et_pb_image][\/et_pb_column][\/et_pb_row][et_pb_row column_structure=”1_5,1_5,1_5,1_5,1_5″ _builder_version=”4.4.7″ min_height=”23px” custom_margin=”|auto|-416px|auto||” custom_padding=”||16px|||”][et_pb_column type=”1_5″ _builder_version=”4.4.7″ custom_padding=”||0px||false|false”][et_pb_text _builder_version=”4.4.7″ header_2_font=”Abhaya Libre|600|||||||” header_2_text_color=”#ffffff” header_2_font_size=”35px” width=”120%” custom_padding=”|49px||0px||”]<\/p>\n

Luca Pizzale<\/h2>\n

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Elodie Wardle<\/h2>\n

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\u00a0John Grissinger<\/h2>\n

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Matt Oatman\u00a0<\/h2>\n

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Katia Krishtopa<\/h2>\n

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Dr. Carl Sparrow<\/span><\/strong><\/h1>\n

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Kif\/Yap 2021\/2022 Project –\u00a0The Effect of KIF11 Activity on YAP LocalizationLuca Pizzale (Form VI), Elodie Wardle (Form VI), \u00a0John Grissinger (Form V), Matt Oatman (Form V), \u00a0Katia Krishtopa (Form IV)Project Summary + Project Goal\u00a0This project is founded upon targeted gene\u00a0therapy and two potential\u00a0proteins\u00a0involved: KIF11 and YAP. KIF11, a motor\u00a0protein, and YAP, a\u00a0transcription factor are both […]<\/p>\n","protected":false},"author":68,"featured_media":0,"parent":0,"menu_order":0,"comment_status":"closed","ping_status":"closed","template":"","meta":{"_et_pb_use_builder":"on","_et_pb_old_content":"","_et_gb_content_width":"","footnotes":""},"class_list":["post-416","page","type-page","status-publish","hentry"],"_links":{"self":[{"href":"https:\/\/students.pingry.org\/irt\/wp-json\/wp\/v2\/pages\/416","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/students.pingry.org\/irt\/wp-json\/wp\/v2\/pages"}],"about":[{"href":"https:\/\/students.pingry.org\/irt\/wp-json\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"https:\/\/students.pingry.org\/irt\/wp-json\/wp\/v2\/users\/68"}],"replies":[{"embeddable":true,"href":"https:\/\/students.pingry.org\/irt\/wp-json\/wp\/v2\/comments?post=416"}],"version-history":[{"count":29,"href":"https:\/\/students.pingry.org\/irt\/wp-json\/wp\/v2\/pages\/416\/revisions"}],"predecessor-version":[{"id":800,"href":"https:\/\/students.pingry.org\/irt\/wp-json\/wp\/v2\/pages\/416\/revisions\/800"}],"wp:attachment":[{"href":"https:\/\/students.pingry.org\/irt\/wp-json\/wp\/v2\/media?parent=416"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}