#Lee Matalone

[janedances]

“It’s impossible to be in love with all the six queens’

The Six queens in question:

Megan Gilbert, Ashleigh Weir, Holly Musgrave, Oliver Wickham, Annabel Marlow, Shimali De Silva, Renee Lamb, Christina Modestou, Natalie Paris, Genesis Lynea, Aimie Atkinson, Izuka Hoyle, Jaye’J Richards-Noel, Millie O’Connell, Alexia McIntosh, Maiya Quansah-Breed, Grace Mouat, Vicki Manser, Courtney Stapleton, Adrianna Hicks, Andrea Macaseat, Abby Mueller, Brittney Mack, Samantha Pauly, Anna Uzele, Mallory Maedke, Nicole Kyoung-Mi Lambert, Courtney Mack, Shantel Cribbs, Courtney Bowman, Sophie Isaacs, Danielle Steers, Zara Macintosh, Cherelle Jay, Hana Stewart, Collette Guitart, Candace Furbert, Hazel Karooma-Brooker, Caitlin Tipping, Sophie Golden, Alicia Corrales-Connor, Viquichele Cross, Bryony Duncan, Natalie Pilkington, Lori McLare, Amy Bridges, Lauren Drew, Maddison Bulleyment, Lauren Byrne, Shekinah McFarlane, Jodie Steele, Athena Collins, Cassandra Lee, Jennifer Caldwell, Harriet Watson, Jasmine Shen, Kelly Sweeney, Jessica Niles, Georgia Carr, Amelia Walker, Liv Alexander, Elizabeth Walker, Maddison Firth, Laura Blair, Chloe Zuel, Kala Gare, Loren Hunter, Kiana Daniele, Courtney Monsma, Vidya Makan, Ella Burns, Karis Oka, Shannen Alyce-Quan, Jade Marvin, Lucy Aiston, Gabriella Stylianou-Burns, Scarlet Gabriel, Rebecca Wickes, Megan Leung, Sophie Rose Middleton, Abbi Hodgson, Kara Ami Mcraenor, Emily Harrigan, Gabrielle Smith, Melissa Ford, Kaylah Attard, Fia Houston-Hamilton, Rhiannon Bacchus, Rhiannon Doyle, Carly Mercedes Dyer, Elena Gyasi, Keirsten Hodgens, Artemis Chrisoulakis, Ellie Sharpe, Sadie Hurst, Melinda Porto, L’Oreal Roache, Wesley Carpenter, Maya Christian, Brianna Mooney, Meghan Dawson, Marilyn Caserta, Ashlee Waldbauer, Adrianna Glover, Alize Ke’Aloha Cruz, Kristina Walz, Amy Di Bartolomeo, Amanda Lindgren, Claudia Kariuki, Dionne Ward-Anderson, Tsemaye Bob-Egbe, Meesha Turner, Paisley Billings, Danielle Rose, Roxanne Couch, Esme Rothero, Rachel Rawlinson, Lauren Irving, Danielle Mendoza, Shelby Griswold, Kennedy Carstens, Abigail Sparrow, Jarynn Whitney, Madeline Fansler, Channing Weir, Princess Victomé, Sunayna Smith, Chloë Hart, Casey Al-Shaqsy, Aiesha Pease, Jaina Brock-Patel, Alana Robinson, Grace Melville, Leesa Tulley, Harriet Caplan-Dean, Khaila Wilcoxon, Storm Lever, Jasmine Forsberg, Olivia Donalson, Didi Romero, Gabriela Carrillo, Cassie Silva, Kelly Denice Taylor, Erin Ramirez, Kelsee Kimmel, Phoenix Mendoza, Chelsea Dawson, Chiara Assetta, Cristina D’Agostino, Joy Woods, Bre Jackson, Keri Rene Fuller, Brennyn Lark, Ayla Ciccone-Burton, Holli’ Conway, Brianna Javis, Gabbi Mack, Casey Esbin, Ellie Wyman, Sasha Renae Brown, Nicole Lamb, Aja Simone Baitey, Willow Dougherty, Kayla McSorely, Emily Rose Lyons, Chelsea Wargo, Hannah Taylor, Jessie Bodner, Jasmine Hackett, Janice Rijssel, Lucia Valentino, Elena Breschi, Meg Dixon-Brasil, Sarah McFarlane, Reca Oakley, Gerianne Perez, Zan Berube, Amina Faye, Terica Marie, Aline Mayagoitia, Sydney Parra, Jana Larell Glover, Taylor Pearlstein, Aryn Bohannon, Cecilia Snow, Rhianne Louise McCaulsky, Baylie Carson, Koko Basigara, Monique Ashe Palmer, Leah Vassell, Hailee Kaleem Wright, Leandra Ellis Gaston, Bella Coppola, Nasia Thomas, Zoe Jensen, Taylor Iman Jones, Aubrey Matalon, Kristina Leopold, Rae Davenport, Gianna Grosso, Kathryn Kilger, Bethany McDonald, Jillian Worthing, Haley Izurieta, Jasmine Smith, Lois Ellise Reeves, Alyssa Giannetti, Eden Holmes, Jaelle Laguerre, Kate Zulauf, Lee ARumSoul, Son Seungyeon Kim Ji Woo, Sophiya Pae, Park Hye-na, Park Ga-Ram, Kim Ji Sun, Choi Hyun-sun, Kim Ryeo Won, Heo Sol-ji, Yoo Ju-hye, Hong Ji Hee, Nicole Louise Lewis, Laura Dawn Pyatt, Erin Caldwell, Kenedy Small, Lou Henry, Aoife Haakenson, Ellie Jane Grant, Izi Maxwell, Tamara Morgan, Shakira Simpson, Fiorella Bamba, Lucinda Wilson, Caitlyn De Kuyper, Amanda Lee, Gabriella Boumford, Audrey Fisher, Brooke Aneece, Jaz Robinson, Julia Pulo, Maggie Lacasse, Krystal Hernández, Elysia Cruz, Lauren Mariasoosay, Julia McLellan, Darcy Stewart, Hailey Lewis

[sixcostumerefs]

Performance Stats: 2022 (mainland edition)

Alright, it’s (no longer) that time of year again! Summary of all 2022 calendar year performances stats!! - This was a CRAZY year full of cross-production covers + emergency cover chaos so very fun to write up. It also made it a bit of a challenge with organization. If an actor was primarily with one production but served as an emergency cover for another, I listed them with their primary or initial production. - Yearly disclaimer that some errors and/or disparities could exist. - And as always, if you repost/use these stats anywhere, please credit me @sixcostumerefs (or six.costume.refs on Insta) And as always....a shoutout to Dionne Ward-Anderson, who featured in the largest number of performances this year, at a whopping 353 shows! Runner up is Andrea Macasaet at 327 shows, with Brittney Mack a very close third with 322! Most performing Aragon was Phoenix Jackson Mendoza (270), Seymour was Claudia Kariuki (293), Howard was Samantha Pauly (314.5), and Parr was Meesha Turner (267.5, but Alana Robinson was at 267). 2021-22 West End Amy di Bartolomeo: 242 performances Amanda Lindgren 255 performances Tsemaye Bob-Egbe: 233 performances Meesha Turner: 267.5 performances Paisley Billings: 114.5 performances (55 A, 44.5 C, 14 P) 2022-23 West End Rhianne-Louise McCaulsky: 81 performances Baylie Carson: 78 performances Koko Basigara: 65 performances Monique Ashe-Palmer: 14 performances (7 A, 1 B, 6 C) Leah Vassell: 40 performances (20 S, 1 C, 19 P) 2021-22 UK Tour Lauren Drew: 71 performances Maddison Bulleyment: 62 performances Caitlin Tipping: 68 performances Shekinah McFarlane: 60 performances Vicki Manser: 63 performances Elena Gyasi: 66 performances (61 regular prior to cast change + 5 emergency cover post-CC) Cassy Lee: 22 performances (8 A, 14 C) Cherelle Jay: 7 performances (1 B, 1 S, 2 C, 3 P) 2022-23 UK Tour Chloe Hart: 269 performances Casey Al-Shaqsy: 252 performances total (249 on UKT + 3 as E/C for WE) Aiesha Pease: 7 performances Jessica Niles: 237-245 performances total (19-27 performances with Breakaway 2.0, 218 performances with UKT) Jaina Brock-Patel: 110 performances Rebecca Wickes: 72 performances Alana M Robinson: 267 performances Harriet Caplan-Dean: 72 performances (10 A, 9 B, 13 S, 7 C, 18 H, 15 P) Grace Melville: 153 performances total w/ 151 performances on UKT (55 A, 4 B, 91 C with 60 of those performances as T/R, 1 P) + 2 as E/C for WE (2 C) Leesa Tulley: 186 performances (1 A, 35 B, 11 S, 137 H with 57 of those performances as T/R, 2 P) 2021-22 Broadway (+ Aug replacements) Adrianna Hicks: 202 performances Bre Jackson: 123 performances (two of these were half shows) Andrea Macasaet: 327 performances Abby Mueller: 117 performances Keri Rene Fuller: 183 performances (59 as T/R, 1 as early debut, the rest as principal) Brittney Mack: 322 performances Samantha Pauly: 314.5 performances Anna Uzele: 97 performances Joy Woods: 146 performances Brennyn Lark: 85 performances Mallory Maedke: 67.5 performances (16.5 A, 47 S, 4 H) Nicole Kyoung-Mi Lambert: 95 performances (39 A, 26 B, 30 C; two A were half-shows) Courtney Mack: 99.5 performances (25 B, 57.5 H, 17 P) Keirsten Hodgens: 107 performances total. 104 on Bway + 3 as emergency cover for Aragon Tour (Bway was 33 S, 25 C, 46 P; Tour was 3 C). Ayla Ciccone-Burton: 20 performances total. 12 pre-CC + 8 post-CC (pre-CC was 2 B, 3 C, 7 P; post-CC was 2 B, 3 C, 3 P) Holli’ Conway: 18 performances total. 9 pre-CC + 9 post-CC (pre-CC 3 C, 6 H; post-CC 5 A, 4 H) Hana Stewart: 2 performances (1 A, 1 P) 2022-23 Broadway Hailee Kaleem Wright: 27 performances Leandra Ellis-Gaston: 30 performances Bella Coppola: 30 performances Nasia Thomas: 29 performances Zoe Jensen: 19 performances Taylor Iman Jones: 29 performances Kristina Leopold: 2 performances (all as S) Aubrey Matalon: 9 performances (all as H) Aragon Tour Khaila Wilcoxon: 258 performances Storm Lever: 267 performances Jasmine Forsberg: 267 performances Olivia Donalson: 263 performances Didi Romero: 261 performances Gabriela Francesca Carrillo: 257 performances Cassie Silva: 41 performances total. 4 with Broadway + 37 with Aragon Tour (Broadway 3 B, 1 H; Aragon Tour 10 B, 13 C, 14 H) Kelly Denice Taylor: 29 performances total. 3 with Boy + 26 with Aragon Tour (Bway 3 S; Aragon Tour was 12 A, 7 S, 7 C) Kelsey Kimmel: 43 performances (16 A, 12 S, 15 P) Erin Ramirez: 34 performances (9 B, 11 H, 14 P) Boleyn Tour Gerianne Perez: 95 performances Zan Berube: 96 performances Amina Faye: 89 performances Terica Marie: 93 performances Aline Mayagoitia: 91 performances Sydney Parra: 96 performances Cecilia Snow: 18 performances (4 A, 11 S, 3 C) Tay Pearlstein: 13 performances (5 B, 7 H, 1 P) Jana Larell Glover: 20 performances (5 A, 8 C, 7 P) Aryn Bohannon: 13 performances (3 B, 4 S, 6 H) Australia Tour Phoenix Jackson Mendoza: 270 performances Kala Gare: 283 performances Loren Hunter: 291 performances Kiana Daniele: 269 performances Chelsea Dawson: 267 performances Video Makan: 254.5 performances Karis Oka: 137 performances (13 A, 28 B, 1 S, 26 C, 38 H, 31 P) Shannen Alyce Quan: 86 performances (9 A, 14 B, 30 S, 2 C, 6 H, 25 P) Chiara Assetta: 93 performances (29.5 A, 3 B, 3 S, 32 C, 18 H, 6.5 P) Cristina D’Agostino: 7 performances (all as P) Madeline Fansler: performances (with Breakaway 3.0: . 16 with Aus Tour: 7 A, 4 S, 5 P) Actors who primarily continued with their initial productions: Claudia Kariuki: 293 performances - 231 pre-CC - 62 post-CC Dionne Ward-Anderson: 353 performances - 269 pre-CC - 84 post-CC Roxanne Couch: 178 performances - Totalling 1 A, 5 B, 61 S, 13 H, 98 P - 122 pre-CC (1 A, 5 B, 61 S, 13 H, 42 P; two of the P were half performances) - 56 post-CC as principal Parr Rachel Rawlinson: 166 performances - Totalling 44 A, 13 B, 51 S, 29 C, 16 H, 13 P - 142 pre-CC (40 A, 12 B, 37 S, 29 C, 13 H, 11 P; 1 C + 1 S were mid-show swing-ons) - 24 post-CC (4 A, 1 B, 14 S, 3 H, 2 P) Esme Rothero: 153 performances - Totalling 19 A, 32 B, 19 S, 16 C, 27 H, 40 P - 129 pre-CC (17 A, 31 B, 19 S, 16 C, 19 H, 27 P) - 24 post-CC (2 A, 1 B, 8 H, 13 P) Danielle Rose: 192 performances - 152 pre-CC (6 A, 51 B, 4 S, 3 C, 87 H, 1 P) - 40 post-CC (15 B, 21 H, 4 P) Jennifer Caldwell: 289 performances total - Totalling 296 as B, 11 H, 3 P - 21 pre-CC as alt (7 B, 11 H, 3 P) - 287 post-CC as principal Boleyn - 2 as E/C Boleyn for West End Natalie Pilkington: 142 performances total - Totalling 8 A, 3 B, 65 S, 12 C, 2 H, 52 P - 130 performances with 22-23 UKT (2 A, 3 B, 64 S, 12 C, 1 H, 48 P) - 3 performances w 21-22 UKT (1 A, 1 H, 1 P) - 9 performances as E/C for WE (5 A, 1 S, 3 P) Standby swings: Harriet Watson: 78 performances total - Totalling 2 A, 19 B, 21 S, 14 C, 6 H, 14 P - 42 performances as alt for the 21-22 UKT (10 B, 11 S, 4 C, 5 H, 12 P) - 3 performances as E/C on the UKT (3 C) - 17 performances as E/C for the WE (6 B, 6 S, 2 C, 1 P) - 6 performances as standby swing for 21-22 WE (2 B, 3 S, 1 H) - 10 performances as standby swing for 22-23 WE (2 A, 1 B, 1 S, 5 C, 1 P) Marilyn Caserta: 5-7 performances total - all as Aragon - 2-4 Bliss 3.0 (all as A) - 3 Bway (all E/C as A) Emergency Covers Courtney Bowman: 1 performance (WE)

[poetsandwriters]

The one activity I’ve found that serves as both social consolation and jolt to my stilted writing is reading Transit and Kudos, the last two books in Rachel Cusk’s Outline trilogy....Cusk’s trilogy offers a particular form of comfort, for this writer at least: No matter what happens—death, divorce, disruption—we have these words that we can wield some control over, when there is little else in this world that we can control.

Lee Matalone, in this week’s Writers Recommend; read the rest at pw.org!

[judgingbooksbycovers]

Home Making

By Lee Matalone.

[bigtickhk]

The Illness Lesson by Clare Beams https://amzn.to/39PUlVp

 Home Making by Lee Matalone https://amzn.to/2HxYz8j

 Vera Violet by Melissa Anne Peterson https://amzn.to/2ueF0ig

[natbrut]

Issue 9 is finally here.

We are super proud of this incredible collection. A lot of work, blood, sweat, and literal tears have gone into this issue. We can't wait for you to read it. Issue 9 features art, edited by Ximena Alejandra Izquierdo Ugaz and Danielle Wright; fiction, edited by RL Goldberg; poetry, edited by Jennifer Soong; and creative nonfiction, edited by Laura Bullard.

Inside you'll also find Another Closet, a folio of work by survivors and members of the LGBTQIA+ community grappling with addiction and substance misuse, edited by Kayla AE and Laura Bullard.

[therumpus]

Some men, some women even, look at us and wonder what we grieve. Let us show you what we now grieve: our belief that our bodies were ours, our belief that we as Americans were above such enmity, that our democracy was above it, that our brothers and mothers and sisters were above it, that progress was a possibility, that we were finally, slowly, getting there, that the days of Styrofoam and plastic bags and back-alley abortions and burnt churches were in the rearview. Grief, in the best cases, brings knowledge, offers a way forward.

R.I.P. #8: Inauguration Day

[yourdailyqueer]

Hey, I was wondering if you could offer any queer YouTube rs to watch other than Mac and Miles. Thank you!

I don’t follow a lot of queer Youtubers but check out the following below. Do note a lot of drag artists from Drag Race are listed because they have Youtube channels now. Be aware these are only the Youtubers that have been posted and not ones in queue or drafts.

Transgender

Charlie Christina Martin

Elena Genevinne

Michelle Hendley

Harrison Browne

Ophelia Pastrana

Miles McKenna

Schuyler Bailar

Emma Ellingsen

Narcissa Wright

Jazz Jennings

Kovu Kingsrod

Gigi Gorgeous

Aaron Ansuini

Elliot Fletcher

Corey Maison 

Angela Vanity

Ryan Cassata

Nicole Ramos

Nikita Dragun

Skylar Kergil

Natalie Wynn

Jamie Raines

Kdin Jenzen

Siri Lehland

Trinity Anne

Laith Ashley

Stef Sanjati

Sam Collins

Kat Blaque

Alex Bertie

Mila Jam

Non binary

Sebastian Columbine

Shiva Raichandani

Kaitlyn Alexander

Chandler Wilson

Brendan Jordan

Thomas Halbert

Madison Paige

Jake Edwards

Annie Segarra

Chris Crocker

Milo Stewart

Jazmin Bean

Ash Hardell

Chella Man

Jude Karda

Kevin Ninh

Dani Shay

B. Scott

Queer/fluid

Cameron Deacon - John Deacon from Queen’s son

Jim Sterling

Daniel Howell

Anna Akana

Rowan Ellis

Jeffery Star

Saintraja

Ari Fitz

Pansexual

Scott Hoying

Ty Turner

Gay

Pedro Álvarez - Spanish speaking

Austin and Aaron Rhodes

The Shirtless Violinist

Nina Bo'nina Brown

Eugene Lee Yang

Zander Hodgson

Lucas Cruikshank

Lasizwe Dambuza

Michael Buckley

Greyson Chance 

Jaymes Mansfield 

Thomas Sanders

Greyson Chance

Raymond Braun

Reuben Mourad

Michael Pavano 

David K. Smith 

Jordan Doww

Jack Merridew

Tuure Boelius

Matthew Lush 

Bretman Rock

Cameron Cole

Nick Crompton

Ryland Adams

Max Emerson

Bilal Hassani

Garrett Watts 

Robert White 

Elijah Daniel

Manny MUA

Matt Dallas

Idan Matalon 

Joey Graceffa

Isaiah Larkin

James Butler

Mitch Grassi

Bryan Odell

Wayne Goss

Trixie Mattel

Allan Alvarez

Troye Sivan

Pabllo Vittar

Tyler Oakley

Kimora Blac

Craig Dillon

Jack Baran 

Tom Daley

Ben Hunte

Phil Lester

Sam Tsui 

Rich Lux

Kingsley

Lesbian

María José Garzón

Shannon Beveridge

Cassandra Bankson

Dr Sally Le Page

Cammie Scott

Christel Dee

Ingrid Nilsen

Hannah Hart

Rose Dix

Hartbeat

Bisexual

Snow Tha Product

Claire Margaret Corlett

Harris “Harry” Brewis

Rosie Spaughton

Andrea Russett

Shane Dawson

Tana Mongeau

Harmony Nice

Janet Devlin

Mikey Bustos

Nicole Pacent

Daniela Calle

Alex Elmslie

Jessie Paege

Sammy Paul

Bree Essrig

Oliver Thorn - Great for philosophy

Lindsay Ellis

Mark Ferris

Gaby Dunn

Meg Turney

Dodie Clark 

Jon Cozart

Lilly Singh

Elle Mills

Mxmtoon

Snooki

Asexual spectrum

Evan Edinger - He reblogged his post

Julie Sondra Decker

Vesper (QueerAsCat)

Connie Glynn

Yasmin Benoit

Caligo Bastet

Ricky Dillon

Intersex

Amythest Schaber

[labdiary]

From the Cultural Revolution to the Gene Therapy RevolutionMay 4, 2020

Guangping Gao, PhD, is professor and director of the Horae Gene Therapy Center at the University of Massachusetts Medical School in Worcester, MA. Over the course of three decades, Gao has made profound contributions in the area of adeno-associated virus research, initially working with James M. Wilson, MD, PhD, director of the gene therapy program at the University of Pennsylvania. Gao has received multiple honors in recognition of his service, expertise, and dedication. For example, he was named president (2019–2020) of the American Society of Gene and Cell Therapy.

Gao has published 250+ research papers, six book chapters, and four edited books, and has fulfilled editorial responsibilities for several gene therapy and virology journals, including the Human Gene Therapy, a journal that Gao currently serves as deputy editor-in-chief. Gao recently spoke to Kevin Davies, PhD, executive editor of Human Gene Therapy, about his remarkable life journey and hopes for the future of gene therapy. (The interview originally appeared in Human Gene Therapy, Vol. 31, Nos. 3 and 4, published by Mary Ann Liebert. Kevin Davies, PhD, executive editor of Human Gene Therapy, conducted the interview.)

We will get to your preeminent research and leadership in the gene therapy field, but let’s start at the beginning.

Gao: I grew up in China during the Cultural Revolution. Around 1975, I was compelled to leave my studies and go to the countryside to receive additional “education” from farmers and peasants. My dream about new medicine really starts there. I interacted with farm laborers on a daily basis, and I saw many of them suffer from various diseases and painful conditions.

I was trying my best to use acupuncture and traditional medicine to help them, but I wished I could have some “magic medicine” to make a more substantial impact, particularly for the elderly and people with cancer.

In 1978, I was one of the first generation of students to enter college after the Cultural Revolution. I was admitted to a medical university in Chengdu, Sichuan. I worked on drug development and medicinal chemistry. In 1988, I graduated from the university and got an opportunity to come to the United States, sponsored by the World Health Organization (WHO). I was looking for opportunities to develop the next generation of medicines that I had dreamed about back on the collective farm.

I started my PhD at Miami Children’s Hospital and Florida International University with my mentor, Reuben Matalon, a pediatrician and medical geneticist. He was a prominent researcher on rare diseases such as Tay-Sachs, Hurler, and Gaucher. His major contribution as a geneticist was the discovery of the biochemical defect in an inherited leukodystrophy called Canavan disease.

I remember it well—I published that paper in the early days of Nature Genetics!

Gao: Yes, thank you! I joined his lab in 1989. My assignment was to isolate the genes and the mutations responsible for Canavan disease. Working with my lab mentor, Rajinder Kaul, I discovered the gene and mutations for Canavan disease and published my thesis work in Nature Genetics in 1993.1

After that, I asked myself, what’s my next step? Because we saw many Canavan patients at these centers, we knew exactly what was going wrong with those kids. We had to figure out a way to fix it. In 1993, I decided to look for the next generation of medicine, specifically at the opportunities in gene therapy for genetic disorders. Finally, Jim Wilson accepted me as a postdoctoral fellow at the University of Pennsylvania’s Institute for Human Gene Therapy.

The first task Jim gave me was to create new generations of adenovirus. At that time, adenovirus vector was much hyped because it has a high transduction efficiency. Because we knew adaptive immunity/immunotoxicity is a major issue for adenovirus, we decided to cripple the virus further to make it more replication defective. This might prolong transduction efficiency and stability in tissues.

I spent about two years there, first making a cell line to complement the crippled virus. Then we used that cell line to create the further-crippled virus. (You need to transcomplement its growth with E1 and E4.) They called this third-generation virus at the time. We demonstrated that, yes, virus can reduce liver toxicity in mice and immunotoxicity and prolong expression substantially.

When I published that work in 1996,2 I said to Jim, “I’d like to move on and start my career in industry because I have two kids to raise.” I was 38 at the time. He said, “No! Why leave? I’m going to give you a job.” He told me they were trying to apply the next-generation adenovirus vector for clinical trials. There was a lab called the Human Applications Lab, a GMP facility at Pennsylvania Hospital where scientists were trying to grow the virus for multiple clinical trials, but they could not grow it well.

My career in gene therapy started from there. I spent about two years making the virus work. In the first two weeks, I was able to generate high quantities of virus. Jim was in his office, talking to a reporter from the Philadelphia Inquirer. I told Jim, “I got the virus, and they are 1013 or 1014.” Jim said to the reporter, “Now we can even swim in this gene therapy vector!”

By that time, we were doing several clinical trials in cystic fibrosis, ornithine transcarbamylase (OTC), mesothelioma, and others. By early 1998, we wanted to look for new viruses, the next generation of gene delivery vehicles. I started working with AAV prototypes such as AAV-2, AAV-1, and AAV-5. Those were the first serotypes to attract a lot of interest and development.

Who first identified AAV? Was it discovered serendipitously?

Gao: Yes, it was discovered in 1965 from some adenovirus preps. They called it adeno-associated virus (AAV) because when they purified the adenovirus and looked at it under a microscope, it was a very small virus in the company of the much larger adenovirus.3 I think Arun Srivastava and others sequenced AAV. Nick Muzyczka, Jude Samulski, Barrie Carter, and others started vectorizing—demonstrating you can create a vector in transduced cells very easily. Many groups then demonstrated that AAV can transduce animals in vivo. The difference is that adenovirus only sustains for a maximum of two to four weeks. But AAV—at that time, primarily AAV-2—can sustain for hundreds of days.

My first task with Jim was to figure out how to produce a scalable manufacturing process. I started making cell lines, creating adeno-AAV hybrids. I published a paper in 1998.4 We converted a transfection-infection system into a total infection system that generates tons of AAV. Working with my colleague Guang Qu, we developed a column purification system using heparin-binding columns in early 2000.

Then on September 17, 1999, this tragic event with adenovirus OTC gene therapy happened, and we lost 19-year-old Jesse Gelsinger. For the entire field, it was a drop from a peak to a deep valley. We experienced 10 years of dark ages for gene therapy. I continued my AAV work. We started the first AAV-2 limb-girdle dystrophy clinical trial with Jerry Mendell (Nationwide Children’s Hospital, Columbus, OH) and colleagues at Penn such as Hansel Stedman and Lee Sweeney. We started the trial using the vector produced with my manufacturing methods under GMP conditions.

After the Gelsinger tragedy, was there added urgency and commitment to establish AAV as an alternative vector?

Gao: Absolutely. We started working with adenovirus, based on the discovery by Yiping Yang (formerly at Duke, now at Ohio State). He discovered immunotoxicity of adenovirus. My job was to reduce that adaptive immunity to adenovirus. But we overlooked this innate immunity, this cytokine storm, which killed Gelsinger.

I had initially started with AAV-2, but we did not really think about AAV-1 and AAV-5, or about discovering new AAVs, until Gelsinger. Then we realized, when you compare the two vectors, adeno is much more efficient. But for immunotoxicity, AAV is much, much better than adeno. Jim and I thought, if we can find a virus as efficient as adeno but without immunotoxicity, that should be the future of gene therapy. Gelsinger was an additional driving force for me to discover new AAVs.

I started work in 2001, and soon we discovered a library of new AAVs in nonhuman primates. We published our first paper in 2002.5 That paper became the hottest paper in the field and gave us new hope to work on the next generation of gene therapy vectors.

How did that discovery come about?

Gao: Back in the winter of 2001, after we found some virus sequences, I presented the PCR data to Jim Wilson at a lab meeting. I could tell his mind was spinning:“Is this real or not?” After the meeting, he said, “Guangping, I think you stepped on a goldmine.”

I started with nonhuman primates. We found that we can detect AAV in any animal. You never run into anyone with absolutely no AAV. It is in any tissue. In any PCR reaction, I always found multiple AAVs. That tells you how diverse [it is], how rapidly AAV is evolving. Then we published our second paper about nonhuman primate viruses, demonstrating AAV evolution.6

At what point did you expand or focus the search for new AAVs in humans?

Gao: You can find AAV everywhere. You can find a different AAV in the same samples. That’s why AAV is amazing to me! As the initial discovery was based on nonhuman primates, I asked Jim in late 2002, “Should we move into human tissues?” He agreed. We discovered AAV-9, which is the first “super virus” for gene therapy from humans, in January 2003.7 Our objective was to develop AAV to be as potent, as efficient, as adenovirus for transduction. But we wanted them to have much less immunogenicity. I think we accomplished that (Figure 1).8

We did not go through the traditional viral isolate characterization. We focused on PCR amplification of the capsid because we realized biology is only determined by the capsid. We didn’t need anything else. We designed PCR primers in the conserved region and amplified through hypervariable regions, generating a new virus capsid with new biology.

When did you move to the University of Massachusetts?

Gao: I moved in 2008. At the time, under the Life Sciences Initiative, then-governor Deval Patrick gave $1 billion to promote biomedicine in the state. Our dean, Terry Flotte, and the chancellor, Michael Collins, wanted to take the momentum to set up three centers in gene therapy, stem cells, and RNA interference. They recruited me from Penn to UMass to set up the gene therapy center.

I continued my AAV discovery, and collaborating with Terry and others—including researchers at the New Iberia (Louisiana) Research Center, a non-human primate facility—we were able to get some primate tissues and start to look for AAV from chimpanzees. We discovered hundreds of AAVs similar to AAV-1, AAV-6, AAV-4, AAV-3, AAV-5, and even AAV-9, which I discovered from humans. I did not know other primates also have AAV-9.

How would you describe the repertoire of AAV vectors? To what degree can researchers adapt these vectors?

Gao: We have now isolated new AAVs from 850 human surgical tissue samples. And we have about 1100 new AAVs. We found large amounts of AAV-2, AAV-3, and AAV-8 in human tissues. My AAV-8 was initially isolated from monkey lymph nodes, but now we see it everywhere in humans. If you talk about the natural reservoir of AAV, I think there is still a lot there.

Of course, now the field has moved to new directions. In addition to a natural reservoir, scientists have started doing directed evolution, rational design, and machine learning. They will complement our original discovery. In the AAV field now, in the clinic, I’d say 98–99% is still the natural AAV as a gene therapy platform, but there are many other AAVs in development by those other methods.

What are the remaining hurdles? Is manufacturing still a challenge?

Gao: If we want to develop clinical AAV gene therapy and commercialize the drugs, we have to overcome four barriers:

Manufacturing. Currently, if you want to use a gene therapy for eyes, for localized delivery to the brain, you don’t need much. Current technology is good enough. But if you want to do things like Duchenne muscular dystrophy or cross the blood-brain barrier, it may require up to 1016 viruses for each patient. In commercial terms, the current maximum scale is probably 1018. But if you are going to use gene therapy and commercialize the drug, usually you need to be on a scale of 1020. We are at least one or two logs away. Generating large quantities of highly potent virus is the number-one barrier we face in the field. This contributes to a major portion of the high cost of gene therapy.

Immunotoxicity. As we are giving AAV at much higher doses, preexisting immunity, innate immunity, and adaptive immunity to capsids and transgenes will become an issue. Some immunotoxicity with high-dose injections is starting to show up. We have to manage this.

Choice. People ask me, “Which AAV do you recommend if I want to target the brain?” That’s a hard question because my understanding, based on natural AAV, is you can either have an efficient or inefficient AAV. There is really a lack of a true tissue tropism, a true cell or tissue specificity. It doesn’t matter how you create a new AAV, that is the area we have to fight for. Eventually we will get there. We’ll make a designer AAV for a certain disease and certain targeted tissue.

Expression. When we do gene therapy, we typically think the more expression, the better. Soon, we will realize that sustained expression at a high, superphysiologic levels may not be good. Particularly with some haploinsufficient diseases, you may run into problems.

References 1. Kaul R, Gao GP, Balamurugan K, Matalon R. Cloning of the human aspartoacylase cDNA and a common missense mutation in Canavan disease. Nat. Genet. 1993; 5: 118–123. 2. Gao GP, Yang Y, Wilson JM. Biology of adenovirus vectors with E1 and E4 deletions for liver-directed gene therapy. J. Virol. 1996; 70: 8934–8943. 3. Hastie E, Samulski RJ. Adeno-associated virus at 50: A golden anniversary of discovery, research, and gene therapy success—A personal perspective. Hum. Gene Ther. 2015; 26: 257–265. 4. Gao GP, Qu G, Faust LZ, et al. High-titer adeno-associated viral vectors from a Rep/Cap cell line and hybrid shuttle virus. Hum. Gene Ther. 1998; 9(16): 2353–2362. 5. Gao GP, Alvira MR, Wang L, et al. Novel adeno-associated viruses from rhesus monkeys as vectors for human gene therapy. Proc. Natl. Acad. Sci. USA 2002; 99: 11854–11859. 6. Gao G, Alvira MR, Somanathan S, et al. Adeno-associated viruses undergo substantial evolution in primates during natural infections. Proc. Natl. Acad. Sci. USA 2003; 100: 6081–6086. 7. Gao G, Vandenberghe LH, Alvira MR, et al. Clades of adeno-associated viruses are widely disseminated in human tissues. J. Virol. 2004; 78: 6381–6388. 8. Wang D, Tai PWL, Gao G. Adeno-associated virus vector as a platform for gene therapy delivery. Nat. Rev. Drug Disc. 2019; 18: 358–378.

[lovelyloveday]

From a talented, powerful new voice in fiction comes a stunning novel about the intersection of three lives coming to grips with identity, family legacy, and what it means to make a house a true home. - Home Making: A Novel by Lee Matalone http://bit.ly/2IFKAxS 

#fiction #contemporary @HarperPerennial #HarperPerennial @Lee_Matalone #LeeMatalone #HomeMaking

[leanpick]

An Adulterer, a Gang Member, a Dystopian Teacher: 3 Novels of American Womanhood

An Adulterer, a Gang Member, a Dystopian Teacher: 3 Novels of American Womanhood

HOME MAKING By Lee Matalone 194 pp. HarperPerennial/HarperCollins. Paper, $15.99.

Image

“Home making” is a phrase so fraught with political and social overtones that before opening this book I went to Google for a sterile definition: “The creation and management of a home, especially as a pleasant place in which to live.” This two-part framework is helpful. While Matalone’s “Home Making” is…

[mironivanov]

An Adulterer, a Gang Member, a Dystopian Teacher: 3 Novels of American Womanhood

Clare Beams’s “The Illness Lesson,” Lee Matalone’s “Home Making” and Melissa Anne Peterson’s “Vera Violet” all star female protagonists at odds with their social surroundings.

[amaised44]

"Objects contain parts of the people who own them.

...

When people go, the things they leave behind can feel haunted. A father can take your left ventricle and a husband can take your frontal lobe, leaving behind a tea kettle, a lampshade from Bed Bath & Beyond. A wooden spoon can hold the existential weight of a family Bible while you turn into a shell."

--Lee Matalone, Home Making

[mittbokligaliv-blog]

Tematrio om starka känslor

Tematrio om starka känslor

Lyrans noblesser frågar: Berätta om tre böcker som innehåller och/eller väcker starka känslor hos dig! 1. Stjärnklart av Lars Wilderäng skrämde skiten ur mig! Den visade hur sårbart vårt samhälle är och att vi är helt beroende av att allt fungerar för att överleva. Bara det här med att få rent vatten är inte helt lätt. 2. Eldens barn av Lucette Matalon Lagnado berörde mig väldigt starkt. Den…

[badgirlnila]

7 Stories About the Anxiety of Settling Down by Lee Matalone via Electric Literature https://ift.tt/37GLeFg

[scvpubliclib]

Clare Beams’s “The Illness Lesson,” Lee Matalone’s “Home Making” and Melissa Anne Peterson’s “Vera Violet” all star female protagonists at odds with their social surroundings.